[Truncated abstract] Hyaline articular cartilage is a highly specialised tissue consisting of chondrocytes embedded in a matrix of proteoglycan and collagens. Hyaline articular cartilage withstands high levels of mechanical stress and continuously renews its extracellular matrix. Despite this durability, mature articular cartilage is vulnerable to injury and disease processes that cause irreparable tissue damage. Native hyaline articular cartilage has poor regenerative capacity following injury, largely due to the tissue's lack of blood and lymphatic supply, as well as the inability of native chondrocytes to migrate through the dense extracellular matrix into the defect site. Articular cartilage injuries that fail to penetrate the subchondral bone plate evoke only a short-lived metabolic and enzymatic response, which fails to provide sufficient new cells or matrix to repair even minimal damage. Clinically, it has previously been accepted that treatment of such defects does not result in the restoration of normal hyaline articular cartilage, which is able to withstand the mechanical demands that are placed on the joint during every day activities of daily living. ... Historically, rehabilitation following ACI has not kept pace with the advances in cell culture and surgical technique. Subsequently, there exists a significant gap in knowledge regarding `best practice' in post operative rehabilitation following ACI. The importance of structured rehabilitation in ACI should not be underestimated when evaluating the clinical success of this chondral treatment. Patients should not be left to their own devices following ACI surgery, as the risk of damage to their implant (via delamination) is high if immediate postoperative movement is not controlled. Furthermore, the biological longevity and clinical success of the graft is dependent on a controlled and graduated return to ambulation and physical activity, and the biomechanical stimulation of the implanted chondrocytes.

A direct bone tendon junction consists of four zones: tendon, uncalcified fibrocartilage, calcified fibrocartilage, and bone. The uncalcified and calcified fibrocartilage together forms the transition zone. This organization ensures a gradual transition in stiffness and material properties, and protects the junction from failure. Transition zone regeneration during bone tendon junction healing is important to restore this unique protective mechanism. / Bone tendon junction repair is involved in many orthopaedic reconstructive procedures. Healing is observed to be slow. The junction often heals by fibrous tissue formation. Previous attempts to enhance bone tendon junction healing have resulted in increased bone formation. However, fibrocartilage transition zone is not restored. / This thesis describes a series of studies on transition zone regeneration in bone tendon junction healing using two partial patellectomy animal models. The healing process inside a bone trough was first studied and characterized. Little transition zone regeneration was observed except near the articular cartilage cut surface. The possibility of using articular cartilage to stimulate transition zone regeneration was explored. Both articular cartilage autograft and allogeneic cultured chondrocyte pellet implantations resulted in significantly increased fibrocartilage transition zone regeneration. Cell tracking indicated that the regenerated tissue likely originated from host cells. To elucidate the mechanism of stimulation by allogeneic cultured chondrocyte pellet, the role of cellular and matrix component needed to be differentiated. Freezing and rapid freeze thaw cycles permanently devitalized the allogeneic cultured chondrocyte pellet, but retained its structural integrity and matrix contents. Preliminary results indicated that implantation of the devitalized allogeneic cultured chondrocyte pellet could still increase fibrocartilage transition zone regeneration. Cellular activity seemed not to be essential for the stimulatory effect. / With further research and development, it is envisioned that a cartilage-based stimulation method for fibrocartilage transition zone regeneration in bone tendon junction healing will be developed for clinical application. / Wong Wan Nar, Margaret. / Source: Dissertation Abstracts International, Volume: 70-06, Section: B, page: 3423. / Thesis (M.D.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 216-231). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

Dissertation submitted in partial compliance with the requirements for the Master’s Degree in Technology: Chiropractic Faculty: Health Sciences, Durban University of Technology, 2009 / Aim To determine the profile of traumatic cervical spine fractures with respect to the epidemiology, clinical presentation, types of fractures, conservative and surgical intervention, short-term post-intervention (i.e. post-conservative and post-surgical) complications and short-term post-surgical rehabilitation of patients presenting at the Spinal Unit of King George V Hospital over a 12-week period. Methods Patients who presented to the King George V Hospital Spinal Unit from surrounding hospitals with traumatic cervical spine fractures were evaluated by the medical staff. Data concerning the epidemiology, clinical presentation, types of fractures, conservative and surgical intervention, short-term post-intervention (i.e. post-conservative and postsurgical) complications and short-term post-surgical rehabilitation data were recorded by the researcher. A p-value of <0.05 was considered as statistically significant. Appropriate statistical tests were applied to the hypothesis-testing objectives. These involved the Pearson’s Chi Square Tests for categorical variables or Fisher’s Exact Tests as appropriate where sample sizes were small. Paired t-tests were done to compare preand- post-surgical Frankel grading and Norton Pressure Sore Assessment scores. Results The number of patients who presented to the Spinal Unit over a 12-week period was 20, of this number 17 were males, three were females and all were black. Eleven patients were treated surgically while nine patients were treated conservatively. The most frequent aetiology of cervical spine fractures was motor vehicle accidents (n = 10) followed by falls (n = 9). The most common co-existing medical conditions were smoking (n = 7), HIV (n = 5), alcohol abuse (n = 3) and obesity (n = 3). The most frequent locations of cervical spine fractures were C2 (n = 6), C1 (n = 4) and the posterior column of C6 (n = 3), while dislocations occurred primarily at the C5-C6 levels (n = 5) of the lower cervical spine. Odontoid fractures (n = 6), Jefferson’s fractures (n = 4) and unilateral facet dislocations (n = 6) were the most common fractures and dislocations v observed. Head injuries (n = 4) and lower limb fractures (n = 3) were the most common extra-spinal fractures. All subjects who sustained head injuries also had associated C1 or C2 fractures. Neurological complications most frequently involved the upper limb where loss of motor function (n = 8) and weakness (n = 4) were observed. The majority of the patients (n = 8) reported a Frankel Grading of E. There were no significant associations between types of fracture and gender with the exception of fracture/dislocation observed in two females. There was a statistically significant difference in the NPSA score (p = 0.004). Conservative care utilized included soft collar (n = 6), cones calipers (n = 6), physiotherapy (n = 4), Minerva jacket (n = 4) and SOMI (sterno-occipital mandibular immobilization) brace (n = 1) while surgical intervention included anterior decompression (n = 8), anterior fusion (n = 8), allograft strut (n = 8), discectomy (n = 8), anterior cervical plating (n = 8), anterior screw fixation (n = 2), a transoral approach (n = 1) and a corpectomy (n = 1). The short-term post-conservative care complications observed in this study were an occipital pressure sore (n = 1), severe discomfort (n = 1) as well as severe neck pain (n = 1), while the short-term post-surgical complications were severe neck pain (n = 2), oral thrush (n = 1), pneumonia (n = 1), odynophagia (n = 1) and hoarseness (n = 1). Of the 11 patients who underwent cervical spine surgery, ten were sent for physiotherapy and one for occupational therapy. No significant associations were seen between the type of cervical spine fracture and the age of the subject. There was a significant association between fracture/dislocation and the female gender (p = 0.016). There was significant negative association between odontoid fracture and: anterior decompression, anterior fusion, allograft strut, discectomy and anterior cervical plating (p = 0.006). Conclusion The results of this study reflect the presentation and management of cervical spine fractures at a referral spinal unit of a public hospital in KwaZulu Natal. The impact of HIV and other co-existing medical conditions were not determined due to the small sample size in this study. Further epidemiological studies are required to be conducted in the Spinal Units of all South African public hospitals in order to confirm or refute the observation of this study. / National Research Foundation (NRF)

Thesis (MMed (Surgical Sciences. Orthopaedic Surgery))--University of Stellenbosch, 2010. / Objective: This research paper reports on the radiographic outcome of unstable thoracolumbar injuries with short segment posterior instrumentation as standalone treatment; in order to review rate of instrumentation failure and identify possible contributing factors. Background: Short segment posterior instrumentation is the treatment method of choice for unstable thoracolumbar injuries in the Acute Spinal Cord Injury Unit (Groote Schuur Hospital). It is considered adequate treatment in fracture cases with an intact posterior longitudinal ligament, and Gaines score below 7 (Parker JW 2000); as well as fracture dislocations, and seatbelt-type injuries (without loss of bone column - bearing integrity). The available body of literature often states instrumentation failure rates of up to 50% (Alanay A 2001, Tezeren G 2005). The same high level of catastrophic hardware failure is not evident in the unit researched. Methods: Sixty-five consecutive patients undergoing the aforementioned surgery were studied. Patients were divided into two main cohorts, namely the “Fracture group” (n=40) consisting of unstable burst fractures and unstable compression fractures; and the “Dislocation group” (n=25) consisting of fracture dislocations and seatbelt-type injuries. The groups reflect similar goals in surgical treatment for the grouped injuries, with reduction in loss of sagittal profile and maintenance thereof being the main aim in the fracture group, appropriately treated with Schantz pin constructs; and maintenance in position only, the goal in the dislocation group, managed with pedicle screw constructs. Data was reviewed in terms of complications, correction of deformity, and subsequent loss of correction with associated instrumentation failure. Secondly, factors influencing the aforementioned were sought, and stratified in terms of relevance. Results: Average follow up was 278 days for the fracture group and 177 days for the dislocation group (all patients included were deemed to have achieved radiological fusion – if fusion technique was employed). There was an average correction in kyphotic deformity of 10.25 degrees. Subsequent loss in sagittal profile averaged 2 degrees (injured level) and 5 degrees (thoracolumbar region) in the combined fracture and dislocation group. The only factor showing a superior trend in loss of reduction achieved was the absence of bone graft (when non-fusion technique was employed). Instrumentation complications occurred in two cases (bent connection rods in a Schantz pin construct with exaggerated loss in regional sagittal profile, and bent Schantz pins). These complications represent a 3.07% hardware failure in total. None of the failures were considered catastrophic. Conclusion: Short segment posterior instrumentation is a safe and effective option in the treatment of unstable thoracolumbar fractures as a standalone measure.

Thesis (MScPhysio)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: Introduction: Iliotibial band syndrome (ITBS), an overuse injury, is the second most common running injury and the main cause of lateral knee pain in runners. Due to the increasing number of runners worldwide there has been an increase in its occurrence. Runners with ITBS typically experience symptoms just after heel strike at approximately 20°-30° of knee flexion (impingement zone) during the stance phase of running. A variety of intrinsic and extrinsic risk factors may be responsible for why some runners are more prone to developing symptoms during the impingement zone as opposed to others. Abnormalities in running biomechanics is an intrinsic risk factor which has been most extensively described in literature but little is known about its exact relationship to ITBS. Objectives: The purpose of this systematic review was to provide an up to date evidence synthesis of the biomechanical risk factors associated with ITBS. These risk factors may need to be considered in the prevention or management of ITBS in runners. A clinical algorithm is also presented. Methods: A systematic review with meta-analysis was conducted. An electronic search was performed in PubMed, PEDro, SPORTSDisc and Scopus of literature published up-until May 2014. Cross-sectional and cohort studies were eligible for inclusion if they evaluated the lower limb biomechanics of runners with ITBS or those who went onto developing it. All studies included in the review were methodologically appraised. Evidence was graded according to the level of evidence, consistency of evidence and the clinical impact. Data was described narratively using tables or narrative summaries where appropriate. A meta-analysis was conducted for biomechanical risk factors which were reported in at least two studies, provided that homogeneity in the outcomes and samples were present. Results: A total of 11 studies were included (1 prospective and 10 cross-sectional). Overall the methodological score of the studies was moderate. Increased peak hip adduction and knee internal rotation during the stance phase may predict the development of ITBS in female runners. These biomechanical risk factors may need to be screened for ITBS prevention, despite the evidence base being limited to a single study. Currently there is no conclusive evidence that any of the biomechanical parameters need to be considered when managing runners with ITBS. Stellenbosch University https://scholar.sun.ac.za iii Conclusion: Biomechanical differences may exist between runners with ITBS and those who may develop ITBS compared to healthy runners. Although a large variety of biomechanical risk factors were evaluated, the evidence base for screening or managing these risk factors for runners with ITBS is limited. This is due to a small evidence base, small clinical effect and heterogeneity between study outcomes and findings. Further prospective and cross-sectional research is required to ascertain if abnormalities in running biomechanics may be related to why runners develop ITBS or to ascertain which risk factors may be involved when managing these runners. / AFRIKAANSE OPSOMMING: Inleiding: Iliotibiale-band-sindroom (ITBS), ’n besering vanweë oormatige gebruik, is die tweede algemeenste hardloopbesering en die hoofoorsaak van laterale kniepyn by hardlopers. Namate die getal hardlopers wêreldwyd toeneem, neem die voorkoms van hierdie toestand ook toe. Hardlopers met ITBS ervaar tipies simptome ná die hakslag met die knie ongeveer 20-30° gebuig (die wrywingsone of “impingement zone”) gedurende die staanfase van hardloop. Verskeie intrinsieke en ekstrinsieke risikofaktore kan ’n rol speel in waarom sommige hardlopers meer geneig is as ander om gedurende die wrywingsone simptome te ervaar. Abnormaliteite in hardloopbiomeganika is ’n intrinsieke risikofaktor wat reeds omvattend in die literatuur beskryf is. Tog is weinig bekend oor presies hoe dit met ITBS verband hou. Oogmerke: Die doel van hierdie stelselmatige ondersoek was om ’n sintese te bied van die jongste bewyse van die biomeganiese risikofaktore van ITBS. Hierdie risikofaktore kan dalk oorweeg word om ITBS by hardlopers te voorkom of te bestuur. ’n Kliniese algoritme word ook aangebied. Metodes: ’n Stelselmatige ondersoek is met behulp van meta-ontleding onderneem. PubMed, PEDro, SPORTSDisc en Scopus is elektronies deurgesoek vir literatuur wat tot en met Mei 2014 verskyn het. Deursnee en kohortstudies is ingesluit indien dit gehandel het oor die biomeganika in die onderste ledemate van hardlopers wat ITBS het of later ontwikkel het. Alle studies wat deel was van die ondersoek is metodologies geëvalueer. Bewyse is aan die hand van bewysvlak, bewyskonsekwentheid en kliniese impak beoordeel. Data is narratief beskryf met behulp van tabelle of narratiewe opsommings waar dit toepaslik was. ’n Meta-ontleding is onderneem waar biomeganiese risikofaktore in minstens twee studies aangemeld is, mits daar homogeniteit in die uitkomste sowel as die steekproewe was. Resultate: Altesaam 11 studies is ingesluit (een prospektief en tien deursnee). Die metodologiese telling van die studies was oorwegend gemiddeld. Verhoogde spitsheupadduksie en interne knierotasie gedurende die staanfase kan op die ontwikkeling van ITBS by vrouehardlopers dui. Hierdie biomeganiese risikofaktore kan dalk nagegaan word vir ITBS-voorkoming, al was die bewysbasis beperk tot ’n enkele studie. Daar is tans geen afdoende bewys dat enige van die biomeganiese parameters oorweeg behoort te word in die bestuur van langafstandatlete met ITBS nie. Gevolgtrekking: Daar bestaan dalk biomeganiese verskille tussen hardlopers wat ITBS het of kan ontwikkel en gesonde hardlopers. Hoewel ’n groot verskeidenheid biomeganiese risikofaktore beoordeel is, is die bewysbasis vir die toets of bestuur daarvan by atlete met ITBS beperk. Dít is vanweë die klein hoeveelheid bewyse, die klein kliniese impak, en heterogeniteit tussen studie-uitkomste en bevindinge. Verdere prospektiewe en deursneenavorsing word vereis om te bepaal of abnormaliteite in hardloopbiomeganika ’n rol kan speel in waarom langafstandhardlopers ITBS ontwikkel, of om vas te stel watter risikofaktore ter sprake kan wees in die bestuur van hierdie hardlopers.

Thesis (MScPhysio)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: This review aims to present the available evidence for the effect of McConnell taping on knee biomechanics in individuals with Anterior Knee Pain (AKP). Pubmed, Medline, Cinahl, Sportdiscus, Pedro and Science Direct electronic databases were searched from inception until September 2014. Experimental research into knee biomechanical or EMG outcomes of McConnell taping compared to no tape or placebo tape were included. Two reviewers completed the searches, selected the full text articles and assessed the risk of bias of eligible studies. Authors were contacted for missing data. Eight heterogeneous studies with a total sample of 220 were included in this review. All of the studies had a moderate to low risk of bias and compared taping to no tape and/ or placebo tape. Pooling of data was possible for three outcomes; average knee extensor moment, average VMO/VL ratio and average VMO-VL onset timing. None of these outcomes revealed significant differences. The evidence is currently insufficient to justify the routine use of the McConnell Taping technique in the treatment of Anterior Knee Pain. There is a need for more evidence on the aetiological pathways of Anterior knee Pain; level one evidence and studies investigating other potential mechanisms of McConnell taping. / AFRIKAANSE OPSOMMING: Die objektief van hierdie resensie was om te bepaal wat die effekte van McConnell Patellar Vasbinding is op knie kinematika, kinetiek en spier aktivering in diegene met Voorafgaande Knie Pyn (VKP). Die navorsers het elektroniese databases soos Pubmed, Medline, Cinahl, Sportdiscus, Pedro en Science Direct, van aanvang tot September 2014, ondersoek. Eksperimenteel studie ontwerpe wat biomeganiese of EMG gevolge van McConnell Vasbinding vergelyk met geen vasbinding of placebo vasbinding, is ingesluit. Twee resente het die ondersoek voltooi, die volle tekse artikels gekies en die partydigheid risiko van die ingeslote studies, geskat. Skrywers is gekontak vir enige verlore data. Agt heterogeen studies uit ‘n totalle monster van 220 is in hierdie resensie ingesluit. Al die studies het ‘n gematigde tot laag risiko vir eensydigheid en vergelyk vasbinding met geen of placebo vasbinding. Data saamvoeging was moontlik vir drie uitslae, naamlik: gemiddelde knie ekstensor moment; gemiddelde VMO/VL ratio en gemiddelde aanval tydmeting. Geen gevolge het veelseggende verskille of afwykings vertoon. Tans is die bewys nie genoegsaam om die routiene gebruik van McConnell Vasbinding tegniek te regverdig nie in die behandeling van VKP. Meer bewyslewering op die etiologiese paaie van VKP; Graad een bewys en studies wat ander moontlike meganisme van Mc Connell Vasbinding ondersoek, is noodsaaklik.

This research focused on three areas of interest regarding feline amputees; owner perception of how their cats adapt to limb amputation, the possibility of phantom sensation, and changes in gait. In general owners felt their cats had an acceptable quality of life; however a proportion believed their cat experienced pain. Anecdotally, owners reported that their cats continued to attempt to use the missing limb following amputation. This prompted the investigation of noninnate forelimb behaviours potentially indicative of phantom sensation; such behaviours apparently persist for months or years after amputation. The impact of phantom sensation on feline welfare is not known, although in humans phantom sensation is a risk factor for phantom pain. Alterations in gait and posture in humans are associated with pain and osteoarthritis. The kinetic changes in feline gait were assessed using a pressure sensitive walkway; this provided its own challenges as the software was designed for large bipeds. As such, a proportion of this work was devoted to developing methods to reliably extract data for small quadrupeds. The results detail how feline amputees alter their weight distribution and paw placement when moving. Observation of amputee cats suggests that they move their limbs differently to those of normal cats, and this was confirmed in a kinematic study using markers to track the motion of each limb. Prior to this research very little was known about how cats coped with limb amputation. The results will better inform the veterinary profession and owners of feline amputees about expected changes, and potentially inform future work on the impact of limb amputation on the welfare of cats. On balance, although there may be some areas of concern, the welfare of cats is acceptable following amputation.

Thesis (Master of Applied Sciences in Chemistry)--Cape Peninsula University of Technology, 2018. / Maintenance of optimum wound chemistry is important to ensure timely healing of a wound. Bacterial infections impair the process of wound healing by producing toxins that alter the chemical environment in and around the wound. The imbalance in the wound chemistry prolongs healing and opens doors to opportunistic infections. Bacteria have developed resistance to conventional bactericides hence, there is need for search of new bactericides that can control bacteria in and around the wound. Therefore, new chemical or biochemical bactericides, which are not resisted by the bacteria, can be explored to control bacterial life around the wound in a bid to maintain optimum wound healing chemistry. Materials such as chitosan, zinc oxide, copper oxide and silver have showed remarkable potential as both bactericidal and wound healing agents. In this work silver, zinc oxide, and copper oxide nanoparticles (NPs) and their chitosan composites (CH-NPs) were synthesized using the chemical reduction method and simple chelation respectively to produce nanoparticles of Ag, ZnO, and CuO as well as composites of CH-ZnO, CH-Ag, CH-CuO, and CH-ZnO-Ag-CuO. Formation of the NPs was confirmed by the exhibition of characteristic peaks in UV-Visible and Fourier Transform Infrared Resonance (FTIR) spectroscopy as well as X-ray diffraction. The nanoparticles (NPs) had optical and electronic band gaps in the range 1 to 5eV indicating their semi-conductive nature. X-ray diffraction (XRD) investigations depicted the crystalline structures of the NPs to be base-centred, face-centred, and hexagonal for Ag, CuO, and ZnO respectively. Transmission electron microscopy (TEM) studies exhibited spherical, hexagonal, and rod-shaped shapes for silver, copper oxide, zinc oxide NPs respectively. Electrochemical investigations of the pure NPs indicated the existence of both the adsorption and the diffusion controlled electron transfer processes at electrode surfaces as well as fast electron transfer rate as depicted by the charge transfer coefficient and standard rate constant parameter values. FTIR spectra of CH-NPs composites depicted new excitation bands absent in spectra of both chitosan and the NPs. The spectra also indicated the deformation and absence of the amine (-NH2) and hydroxyl bands (-OH) within the CH-NPs composites. UV-Visible spectroscopy investigations of the CH-NPs composites exhibited blue-shifts of the λmax with respect to the NPs. The FTIR and UV-Visible spectra confirmed the existence of bonding between the chitosan and the NPs. The optical band gap energies of all the CH-NPs composites fell within the range of 2.0 to 4.5 eV indicating that the CH-NPs fell in the category of the semi-conducting materials after chelating with the chitosan.

肌腱損傷發生率高，並且癒合結果很不理想，因為少量的肌腱細胞缺乏有效的修復能力，僅僅通過瘢痕形成來癒合, 肌腱瘢痕癒合難以恢復原本的肌腱組織結構及力學特性。目前，國內外臨床上治療肌腱損傷的方法很多，包括藥物、物理治療、手術等，這些並不能獲得滿意的療效。因此，如何採用肌腱組織工程技術迅速、安全、有效的修復肌腱損傷已成為運動醫學領域急需解決的重要問題。 / 有研究表明，骨髓間充質幹細胞、表皮成纖維細胞、肌腱細胞和胚胎幹細胞通過肌腱組織工程技術用於肌腱修復及再生取得了不錯的療效。但是，這些來源的細胞存在分化效率低，形成畸胎瘤和異位骨化等風險。近來，有研究報導可從人、小鼠、大鼠和兔的肌腱組織中分離培養出幹細胞，可作為肌腱組織工程種子細胞的一種新選擇，用於肌腱修復和再生。對於間充質幹細胞的成肌腱分化，有研究報導結締組織生長因子（CTGF）和抗壞血酸（維生素C的一種形式）在膠原及細胞外基質合成、調節細胞成肌腱分化方面扮演者重要的角色。 / 本研究的旨在：（1）在大鼠髕腱損傷模型中，證實肌腱幹細胞可作為一種新的幹細胞來源用於肌腱修復；（2）檢驗結締組織生長因子和抗壞血酸能在體外促進肌腱幹細胞的成肌腱分化；（3）嘗試通過肌腱幹細胞的成肌腱分化過程在體外構建不含外源性支架的肌腱樣組織；（4）探索該肌腱樣組織在大鼠髕腱損傷模型中是否可以促進肌腱癒合。 / 在大鼠急性髕腱損傷動物模型中，與對照組相比，肌腱幹細胞組具有更好的膠原排列，顯著增高的最大張力和楊氏模量，表明肌腱幹細胞可作為一種新的幹細胞來源用於肌腱損傷的修復。結締組織生長因子和抗壞血酸體外誘導肌腱幹細胞2周後，可顯著增加Tenomodulin, Scleraxis, Thbs4, I型膠原等肌腱相關基因的表達以及膠原蛋白的合成，說明結締組織生長因子和抗壞血酸可促進肌腱幹細胞的成肌腱分化。被結締組織生長因子和抗壞血酸誘導兩周後，肌腱幹細胞可形成了細胞膜樣結構，將這種細胞膜纏繞在迴紋針上，構建成肌腱樣組織，其具有相對疏鬆的細胞外基質和雜亂排列其中的肌腱幹細胞，以及表達Tenomodulin，I型膠原和III型膠原。將該肌腱樣組織移植到裸鼠體內8周和12周可形成新生肌腱組織，梭形細胞縱行分佈在平行的膠原纖維之間，並表達Tenomodulin，I型膠原和III型膠原蛋白。在大鼠髕腱損傷動物模型中，與對照組相比較，該肌腱樣組織可通過恢復肌腱組織結構及生物力學特性來促進肌腱癒合。 / 總的來說，本研究證實肌腱幹細胞可作為一種新的幹細胞來源用於肌腱組織工程促進肌腱再生。結締組織生長因子和抗壞血酸可調控肌腱幹細胞的成肌腱分化，並形成細胞膜結構。該細胞膜結構可在體外構建出不含外源性支架的肌腱樣組織，進而在裸鼠體內形成新生肌腱，並且在大鼠髕腱損傷模型中可有效的促進損傷肌腱的癒合。這種不含外源性支架的肌腱樣組織有希望成為肌腱組織工程技術的新手段，在肌腱再生和肌腱修復的臨床應用及基礎研究方面有廣泛的前景。 / Tendon injuries are common and tendon healing outcome is poor, because tendon contains few cells with limited capacities for self-repair/regeneration. The current treatments on tendon injuries including drugs, physiotherapy, and surgery are not ideal and there is a need for the development of novel tissue-engineering strategies for tendon repair. / Previous studies have shown positive effects of bone marrow-derived mesenchymal stem cells (BMSCs), dermal fibroblast, tenocytes, and embryonic stem cells-derived MSCs for tendon repair/regeneration. However, these cells have limitations including insufficient differentiation; risk of teratoma and ectopic bone formation etc. Recently, stem cells have been isolated from tendons of human, mouse, rat and rabbit and considered as a new alternative cell source for tendon tissue engineering (TDSCs). For tenogenic differention of MSCs, connective tissue growth factor (CTGF) and ascorbic acid (one form of vitamin C) are reported to play important roles in promoting collagen and other extracellular matrixes (ECM) production, and regulating the MSCs differentiation towards tenogenic pathway. / The aims of the current study are: (1) To investigate the use of TDSCs in tendon repair in a rat acute patellar tendon injury model; (2) To test the effects of CTGF and ascorbic acid on tenogenic differentiation of TDSCs in vitro; (3) To construct scaffold-free tendon-like tissues in vitro using tenogenically differentiated TDSCs; (4) To promote tendon healing by engineered tendon-like tissues in a rat acute patellar tendon injury model. / In the rat acute patellar tendon injury model, in contract to control group, TDSCs treated group showed better alignment of collagen fibers and the significantly higher ultimate stress and Young’s modulus, indicating TDSCs may be an alternative cell source for tendon repair. The effects of CTGF and ascorbic acid on tenogenic differentiation of TDSCs were also confirmed with higher expression of tendon related markers such as Tenomodulin, Scleraxis, Thbs4, Type I Collagen, etc; with higher production of collagenous proteins. After treatment with CTGF and ascorbic acid for 2 weeks, TDSCs can form cell sheets, which can be harvested, rolled up on a U-shaped spring to form tendon-like tissues in culture, which had loose extracellular matrices and randomly distributed TDSCs and also expressed Tenomodulin, Type I & III collagen. Following transplantation of the engineered tendon-like tissue in nude mice for 8 and 12 weeks, neo-tendon tissues were formed, with thin and parallel collagen fibrils and extracellular matrices of Tenomodulin, Type I & III collagen. Finally in the rat patellar tendon window injury model, data suggested that the engineered tendon-like tissue could promote tendon healing with significantly improved histological features and biomechanical properties comparing to the control group. / In conclusion, our study has indicated that TDSCs can be an alternative cell source in tendon tissue engineering for tendon regeneration. The tenogenic differentiation of TDSCs, induced by CTGF and ascorbic acid in vitro, produces cell sheets, which can be constructed tendon-like tissues in vitro; to form neo-tendon and repair tendon injuries in vivo. The use of engineered scaffold-free tendon tissue for tendon tissue engineering has potentials in clinical application for tendon repair/regeneration. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Ni, Ming. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 107-126). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / DEDICATION --- p.I / ACKNOWLEDGEMENT --- p.II-III / TABLE OF CONTENTS --- p.IV-IX / PUBLICATIONS --- p.X-XII / ABBREVIATION --- p.XIII-XV / ABSTRACT (ENGLISH) --- p.XVI-XVIII / ABSTRACT (CHINESE) --- p.XIX-XX / Chapter CHAPTER 1 --- Introduction --- p.1 / Chapter 1.1 --- Epidemiology of tendon injury --- p.1 / Chapter 1.2 --- Healing process of tendon injury --- p.1 / Chapter 1.3 --- Tendon tissue engineering for tendon repair --- p.2 / Chapter 1.4 --- Stem cells in tendon repair --- p.2 / Chapter 1.5 --- Tenogenic differentiation of tendon derived stem cells --- p.7 / Chapter 1.6 --- Growth factors for tenogenic differentiation --- p.8 / Chapter 1.7 --- Vitamin C for tenogenic differentiation --- p.9 / Chapter 1.8 --- Summary --- p.10 / Chapter CHAPTER 2 --- Hypothesis, Objectives and Study Design --- p.11 / Chapter 2.1 --- Hypothesis --- p.11 / Chapter 2.1.1 --- Overall hypothesis --- p.11 / Chapter 2.1.2 --- Specific hypothesis --- p.11 / Chapter 2.2 --- Objectives --- p.12 / Chapter 2.3 --- Study design --- p.12 / Chapter 2.3.1 --- Study I --- p.12 / Chapter 2.3.2 --- Study II --- p.14 / Chapter 2.3.3 --- Study III --- p.14 / Chapter 2.3.4 --- Study IV --- p.17 / Chapter CHAPTER 3 --- Tendon-derived Stem Cells (TDSCs): A New Cell Source for Tendon Repair (Study I) --- p.19 / Chapter 3.1 --- Materials and Methods --- p.19 / Chapter 3.1.1 --- Isolation and characterization of rat GFP-TDSCs --- p.19 / Chapter 3.1.2 --- Animal surgery --- p.20 / Chapter 3.1.3 --- Ultrasound imaging --- p.25 / Chapter 3.1.4 --- Histology --- p.27 / Chapter 3.1.5 --- Biomechanical test --- p.27 / Chapter 3.1.6 --- Ex vivo fluorescence imaging --- p.28 / Chapter 3.1.7 --- Data analysis --- p.29 / Chapter 3.2 --- Results --- p.29 / Chapter 3.2.1 --- Gross observation of the injured knee and patellar tendon --- p.29 / Chapter 3.2.2 --- Histology of regenerated tendon tissue --- p.30 / Chapter 3.2.3 --- Biomechanical test of regenerated tendon tissue --- p.32 / Chapter 3.2.4 --- Ex vivo fluorescence imaging of GFP-TDSCs --- p.33 / Chapter 3.2.5 --- Ultrasound imaging of wound gap volume --- p.34 / Chapter 3.3 --- Discussion --- p.35 / Chapter 3.4 --- Conclusion --- p.50 / Chapter CHAPTER 4 --- Tenogenic Differentiation of Tendon-derived Stem Cells (TDSCs) (Study II) --- p.51 / Chapter 4.1 --- Materials and Methods --- p.51 / Chapter 4.1.1 --- Tenogenic differentiation of tendon-derived stem cells (TDSCs) --- p.51 / Chapter 4.1.2 --- Quantification of collagenous proteins --- p.51 / Chapter 4.1.3 --- Quantitative Real Time PCR (qRT-PCR) --- p.52 / Chapter 4.1.4 --- Data analysis --- p.54 / Chapter 4.2 --- Results --- p.55 / Chapter 4.2.1 --- Quantification of collagenous proteins --- p.55 / Chapter 4.2.2 --- Tenogenic, osteogenic and chondrogenic markers mRNA expression --- p.57 / Chapter 4.2.3 --- Tendon extracellular matrix markers mRNA expression --- p.57 / Chapter 4.3 --- Discussion --- p.59 / Chapter 4.4 --- Conclusion --- p.66 / Chapter CHAPTER 5 --- Engineered Scaffold-free Tendon Tissue Produced by Tendon-derived Stem Cells (TDSCs) Cell Sheet (Study III) --- p.67 / Chapter 5.1 --- Materials and Methods --- p.67 / Chapter 5.1.1 --- In vitro engineered scaffold-free tendon tissue by TDSCs cell sheet --- p.67 / Chapter 5.1.2 --- In vivo neo-tendon formation using engineered scaffold-free tendon tissue in nude mouse model --- p.67 / Chapter 5.1.3 --- Histology and immunohistochemistry staining --- p.68 / Chapter 5.1.4 --- In vivo fluorescence imaging --- p.69 / Chapter 5.1.5 --- Data analysis --- p.70 / Chapter 5.2 --- Results --- p.70 / Chapter 5.2.1 --- Gross observation of TDSCs cell sheet and engineered scaffold-free tendon tissue --- p.70 / Chapter 5.2.2 --- Histological and immunohistochemical characteristics in engineered scaffold-free tendon tissue --- p.71 / Chapter 5.2.3 --- Gross observation and in vivo fluorescence imaging of neo-tendon tissue --- p.74 / Chapter 5.2.4 --- Histology of neo-tendon tissue --- p.75 / Chapter 5.2.5 --- Immunohistochemistry staining in neo-tendon tissue --- p.76 / Chapter 5.3 --- Discussion --- p.78 / Chapter 5.4 --- Conclusion --- p.82 / Chapter CHAPTER 6 --- Use of Engineered Scaffold-free Tendon Tissue for Tendon Repair (Study IV) --- p.83 / Chapter 6.1 --- Materials and methods --- p.83 / Chapter 6.1.1 --- Animal surgery --- p.83 / Chapter 6.1.2 --- Ex vivo fluorescence imaging --- p.84 / Chapter 6.1.3 --- Histology and immunohistochemistry staining --- p.85 / Chapter 6.1.4 --- Biomechanical test --- p.86 / Chapter 6.1.5 --- Ultrasound imaging --- p.87 / Chapter 6.1.6 --- Data Analysis --- p.87 / Chapter 6.2 --- Results --- p.88 / Chapter 6.2.1 --- Gross observation of the injured knee and patellar tendon --- p.88 / Chapter 6.2.2 --- Histology of regenerated tendon tissue --- p.89 / Chapter 6.2.3 --- Tendon specific and ECM markers expression in regenerated tendon tissue --- p.91 / Chapter 6.2.4 --- Osteogenic and chondrogenic specific markers expression in neo-tendon tissue --- p.93 / Chapter 6.2.5 --- The fate of the transplanted engineered scaffold-free tendon tissue --- p.93 / Chapter 6.2.6 --- Biomechanical test of regenerated tendon tissues --- p.94 / Chapter 6.3 --- Discussion --- p.96 / Chapter 6.4 --- Conclusion --- p.102 / Chapter CHAPTER 7 --- General Conclusions --- p.103 / Chapter 7.1 --- General discussion --- p.103 / Chapter 7.2 --- General conclusions --- p.105 / FUNDING --- p.106 / REFERENCES --- p.107 / APPENDIX --- p.127

Traumatic brain injury (TBI) is the signature injury of the ongoing military conflicts in the Middle East and Afghanistan, largely due to the use of improvised explosive devices (IEDs), which have affected soldiers and civilians alike. Blast-induced TBI (bTBI) biomechanics are complex and multiphasic. While research has clearly demonstrated the negative effects of penetrative (secondary blast) and inertia-driven (tertiary blast) injury, the effect of shock wave loading (primary blast) on the brain remains unclear. Combined primary-tertiary blast exposure in vivo has been reported previously to alter brain function, specifically hippocampal function; however, it is extremely difficult to deliver primary blast exposure in isolation with an in vivo injury model. The research presented in this thesis utilized a custom-designed in vitro blast injury model to deliver military-relevant shock wave exposures, in isolation, to organotypic hippocampal slice cultures (OHSCs). To contextualize blast-induced pathobiology with previous TBI studies, the first goal of this thesis was to experimentally characterize the deformation profile induced in OHSCs with our blast injury model. Using stereoscopic, high-speed cameras and digital image correlation to calculate strain, we found that our blast model induced low strain magnitudes (<9%) but at high strain rates (25-86s-1), which aligned closely with associated computational simulations of our model. The second aim was to determine if primary blast was capable of altering hippocampal electrophysiological function. We exposed OHSCs to a range of shock intensities and found, using a micro-electrode array system, that long-term potentiation (LTP), a measure of synaptic plasticity, was very sensitive to primary blast exposure; a threshold for disruption of LTP was found between 9 and 39 kPa•ms impulse. Alternative measures of basal electrophysiology were less sensitive than LTP. Blast exposure significantly reduced LTP between 1 and 24 hours post-injury, and this deficit persisted through 6 days post-injury. Depending on shock intensity, LTP spontaneously recovered 10 days post-injury. The third aim was to explore the cellular mechanisms for blast-induced LTP deficits. Using a chemical LTP induction protocol, blast exposure altered key proteins necessary for the induction of LTP by 24 hours post-injury including, postsynaptic density protein-95 (PSD-95), a major scaffolding protein that organizes the postsynaptic density (PSD), α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid glutamate receptor 1 (AMPA-GluR1), and stargazin, an auxiliary GluR1 protein that binds AMPA-GluR1 to PSD-95. Modulation of the cyclic adenosine monophosphate (cAMP) pathway reversed the observed effects of blast on LTP. We theorized that blast-induced disruption of PSD-95 prevented translocation, and subsequent phosphorylation, of GluR1-containing AMPARs to the postsynaptic membrane, which, in turn, prevented potentiation. The final aim was to investigate the efficacy of phosphodiesterase-4 (PDE4) inhibitors, which block degradation of cAMP, as a therapeutic strategy. When delivered immediately following primary blast injury, multiple PDE4 inhibitors proved efficacious in restoring LTP measured 24 hours post-injury. Roflumilast, a Food and Drug Administration-approved PDE4 inhibitor, was effective when delivered at a clinically relevant concentration (1nM) and at a delayed time point (up to 6 hours). Roflumilast reversed blast-induced changes in expression/phosphorylation of the key LTP protein targets. We hypothesized that maintenance of PSD-95 drove the observed therapeutic effect. Greater work is necessary to determine how blast exposure degrades PSD-95 and how roflumilast prevented these detrimental effects. This thesis has shown that primary blast exposure can negatively alter neurological function, as well as protein expression and phosphorylation. These studies expand the understanding of primary blast injury mechanisms, provide computational models with important tissue-level tolerance criteria, inform protective equipment design, inform clinical care guidelines for bTBI, and present a promising therapeutic candidate for further clinical investigation.