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No. of bitstreams: 2 Dissertação - Alexandre Lamaro Cardoso - 2015.pdf: 3044296 bytes, checksum: a1b0257c1b7a1f083429fbd5076a9887 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2015-05-22 / Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG / Stromal vascular fraction and irradiation with low level laser are resources potencially applicable in the treatment of burn injuries. Objectives This study aimed to: (i) analyze the macroscopic, microscopic and morphometric effects of stromal vascular fraction application compared to conventional care in 3rd degree burn injuries in rats; (ii) analyze the macroscopic, microscopic and morphometric effects of stromal vascular fraction containing adipose derived stem cells application associated with low level laser compared to conventional care in 3rd degree burn injuries in rats; (iii) whether the macroscopic, microscopic and morphometric effects of stromal vascular fraction containing adipose derived stem cells application associated with low level laser irradiation are dependent on the irradiation dose applied. Method: This project was approved by the Ethics Committee on the use of animals at UFG, protocol number 035/2013. All animals received injections of stromal vascular fraction prepared from adipose tissue extracted from a animal donor and occlusive dressing. The animals were divided into 5 groups: control (CT), stromal vascular fraction (SVF), stromal vascular fraction plus Laser at 27mW of power (SVFL27), stromal vascular fraction plus Laser at 53mW of power (SVFL53) and stromal vascular fraction plus Laser at 76mW of power (SVFL76).The animals were followed ove days 3, 7 and 30 for the evaluation of macroscopic, microscopic and morphometric aspects. For statistical analysis we used the parametric "t test". Results: The morphometric analysis showed a reduction in wound area in SVFL27 group compared to the others, but without statistically significant difference. In SVFL76 group was observed inhibition of wound contraction compared to SVFL27 and CT groups (p <0,05). Was observed on day 3 significantly increased (p <0.05) infiltration of polymorphonuclear, fibrin formation and in the number of fibroblasts in all treated groups compared to CT. On the 7th day the mononuclear infiltrate, angiogenesis, collagen and number of fibroblasts was significantly higher in all treated groups compared to CT (p <0.05). The SVFL27 group also showed a higher number of fibroblasts compared to groups SVF, SVFL53 and SVFL76. At 30 days significantly increased reepitalization and collagen deposition were observed in all treated groups compared to CT (p <0.05). The SVFL53 group also showed higher polymorphonuclear infiltrate compared to other groups (p <0.05). Conclusion: Isolated application of SFV or in combination with low level Laser gives better results than conventional care in burn injury in rats and the effects of the combination of the two features are dose / Fração estromal vascular e irradiação com Laser de baixa potência são recursos potencialmente aplicáveis no tratamento de lesões por queimadura. O presente estudo teve por objetivo: (i) analisar os efeitos macroscópicos, microscópicos e morfométricos da aplicação de fração estromal vascular contendo células-tronco derivadas de tecido adiposo comparada ao cuidado convencional em lesões por queimadura de 3º grau em ratos; (ii) analisar os efeitos macroscópicos, microscópicos e morfométricos da aplicação de fração estromal vascular contendo células-tronco derivadas de tecido adiposo associada ao Laser de baixa potência comparada ao cuidado convencional em lesões por queimadura de 3º grau em ratos; (iii) analisar se os efeitos macroscópicos, microscópicos e morfométricos da aplicação de fração estromal vascular associada ao Laser de baixa potência são dependentes da dose de irradiação aplicada. Método: Este projeto foi aprovado pelo Comitê de Ética no uso de animais da UFG, protocolo número 035/2013. Todos os animais receberam injeções de fração estromal vascular preparada a partir do tecido adiposo extraído de animal doador e curativo oclusivo. Os animais foram distribuídos em 5 grupos: controle (CT), fração estromal vascular (FEV), fração estromal vascular mais Laser a 27mW de potência (FEVL27), fração estromal vascular mais Laser a 53mW de potência (FEVL53) e fração estromal vascular mais Laser a 76mW de potência (FEVL76). Foram acompanhados ao longo dos dias 3, 7 e trinta para a avaliação dos aspectos macroscópicos, microscópicos e morfométricos. Para análise estatística foi utilizado o teste Kruskal-Wallis. Resultados: a análise morfométrica mostrou redução da área da ferida mais acentuada no grupo FEVL27 comparado ao demais, porém, sem diferença estatisticamente significante. No grupo FEVL76 foi observada inibição da contração da ferida comparada aos grupos FEVL27 e CT. Foi observado no 3º dia aumento significativo (p <0,05) do infiltrado polimorfonuclear, formação de fibrina e número de fibroblastos em todos os grupos tratados comparados ao CT. No 7º dia o infiltrado mononuclear, angiogênese, colágeno e número de fibroblastos foi significativamente maior em todos os grupos tratados comparados ao CT (p <0,05). O grupo FEVL27 também apresentou maior número de fibroblastos comparado aos grupos FEV, FEVL53 e FEVL76. Aos 30 dias aumento significativo da reepitalização e deposição de colágeno foram observados em todos os grupos tratados comparados ao CT (p <0,05). O grupo FEVL53 também apresentou maior infiltrado polimorfonuclear comparado aos demais grupos (p <0,05). Conclusão: aplicação isolada de FEV ou em associação com Laser de baixa potência confere melhores resultados que o cuidado convencional em lesões por queimadura em ratos e os efeitos da associação dos dois recursos são dose dependentes.

A criocirurgia tem sido utilizada no tratamento de diferentes enfermidades de sistemas e órgãos, tanto na medicina humana quanto na medicina veterinária, sendo sua maior indicação o tratamento de dermatopatias. Contudo, efeitos adversos como a cicatrização lenta, cicatrizes extensas, disfunção estética e funcional, são relatados após a aplicação da substância criogênica. Ainda, existem as lesões que ocorrem naturalmente pela exposição ao frio extremo, afetando principalmente o nariz, dedos das mãos e pés ou orelhas. Na maioria das vezes resultam em gangrena e são bem comuns nos habitantes dos polos, turistas e praticantes de modalidades na neve. O presente trabalho tem como objetivo avaliar a influência das células-tronco mesenquimais de origem adiposa (ADSCs) na cicatrização de feridas cutâneas padronizadas e induzidas pelo nitrogênio líquido em ratos em duas fases da cicatrização cutânea: fase de proliferação e fase de remodelação. Utilizaram-se 83 ratos Wistar, machos, hígidos, com oito semanas de idade, sendo três animais usados como doadores de tecido adiposo para posterior obtenção das ADSCs e, 80 animais divididos aleatoriamente em oito grupos de tratamento (n=10). Através da aplicação do nitrogênio líquido pela técnica do spray aberto, realizou-se a indução de uma ferida, de aproximadamente 15 mm de diâmetro, na região dorsal de cada rato. A ferida recebeu o tratamento de acordo com o grupo ao qual pertencia: a) aplicação das ADSCs, por via subcutânea, no 15º dia (T1), no 30º dia (T2) ou nos dois tempos mencionados (T3) após a indução da lesão; b) aplicação da solução cloreto de sódio 0,9%, por via subcutânea, no 15º dia (S1), no 30º dia (S2) ou nesses dois tempos (S3) após a indução da lesão; c) nenhuma intervenção até o momento da eutanásia dos animais no 45º dia (C1) ou no 60º dia (C2). Macroscopicamente, a cada cinco dias, analisaram-se as medidas das lesões e calculou- se a área e a taxa de contração cicatricial das mesmas. No 45º ou no 60º dia pós- operatório, procedeu-se à coleta das biópsias para avaliação histopatológica e imuno- histoquímica. Com base nos resultados obtidos concluiu-se que: 1) o grupo T1 apresenta as maiores taxas de contração média das feridas no 20º e 25º dia; 2) o grupo T3 obteve a maior taxa de contração média das feridas no 30º dia pós-operatório; 3) o grupo T2 apresenta as maiores taxas de contração média das feridas no 55º e 60º dia; 4) o grupo T1 obteve diferença estatisticamente significativa em relação ao grupo sham (S3) quanto à neovascularização, avaliada pela técnica de imuno-histoquímica com o VEGF; 5) o grupo sham (S1) obteve diferença estatística significativa em relação aos grupos tratados com as ADSCs (T2 e T3) quanto à proliferação epitelial, avaliada pela técnica de imuno-histoquímica com o anticorpo Ki-67; 6) a terapia com as ADSCs proporciona uma relevante evolução clínica das feridas, podendo ser constatada ao final do período de avaliação por cicatrizes mais estreitas e compridas com as medidas da área final inferiores às cicatrizes dos grupos controle (C1 e C2) e sham (S1, S2 e S3). Propõem-se a necessidade de novos estudos com as ADSCs na cicatrização de lesões cutâneas provocadas pela criocirurgia ou por outra modalidade de congelamento, realizando biópsias com análises histopatológicas e imuno-histoquímicas em períodos de tempo menores e maiores aos realizados nesse estudo, a fim de detectar, respectivamente, diferenças no processo de cicatrização imediatamente após a aplicação das ADSCs e, também, acompanhar o remodelamento da cicatriz por um período mais longo. / Cryosurgery has been used to treat different diseases of systems and organs in both human and veterinary medicine. Although treatment of skin disorders is the leading indication, adverse effects such as delayed wound healing, large scars, esthetical deformation and functional impairment have been reported from administration of cryogenic substance. Beside that there are injuries caused naturally by the exposure to extreme cold weather conditions, which affect specially nose, fingers, toes and ears, mostly resulting in gangrene. These frostbites are very common in people who live in the Poles, tourists and snowboarders. This study aims to evaluate the influence of adipose-derived stem cells (ADSCs) on cutaneous wound healing that were standardized and induced by liquid nitrogen in rats according to two phases of cutaneous healing: proliferation phase and remodelling phase. For research purposes, 83 male, healthy and eight-weeks-old Wistar rats were required. Among 83 rats, three were used as adipose tissue donor for later ADSCs obtention and 80 Wistar rats were randomly divided in eight treatment groups (n=10). Through the application of liquid nitrogen by spraying technique, a 15 millimetres in diameter lesion was produced in the dorsal region of each rat. The wound received treatment according to the group it belonged: a) subcutaneously ADSCs application on the 15th day (T1), on the 30th day (T2) or in both periods mentioned (T3) after wound induction; b) subcutaneously application of 0.9% sodium chloride solution on the 15th day (S1), on the 30th day (S2) or in both periods mentioned (S3) after wound induction; c) no intervention until euthanasia on the 45th day (C1) or 60th day (C2) Macroscopically, every five days, the wounds were measured to calculate their area and healing rate. On the 45th and 60th postoperative day, biopsies were performed for histopathological and immunohistochemical evaluations. By the obtained results, the study concludes that:1) T1 group shows the highest wound average contraction rate on the 20th and 25th day; 2) T3 group presents the highest wound average contraction rate on the 30th postoperative day; 3) T2 group has the highest wound average contraction rate on the 55th and 60th day; 4) T1 group got a significant statistical difference in relation to sham group (S3) when it refers to neovascularization, which was evaluated by immunohistochemical technique with VEGF; 5) sham group (S1) obtained a significant statistical difference when compared to ADSCs groups (T2 and T3) with respect to epithelial proliferation, that was evaluated by immunohistochemical technique using antibody Ki-67; 6) the ADSCs therapy provides an important clinical evolution of wounds. This was verified at the end of evaluation period through narrower and longer scars with bottom end area measurements inferior to control group scars (C1 and C2) and sham group scars (S1, S2 and S3). Lastly, this paper proposes the necessity of new studies about the uses of ADSCs for cutaneous wound healing caused by cryosurgery or other sort of freeze. Furthermore it is opportune taking an action on studies that do biopsies with histopathological and immunohistochemical analyses using shorter and longer periods of time that those executed on this paper. Thus it will be possible to find out, respectively, differences on the healing process immediately after applying ADSCs and also follow up the scar remodelling for a longer period.

Degenerative Veränderungen der Lendenwirbelsäule beginnen bereits im Alter von unter 20 Jahren und betreffen vor allem die unteren 3 Bewegungssegmente. Die degenerativen Veränderungen an der Bandscheibe gehen mit einer Kalzifizierung der Grund- und Deckplatten der Wirbelkörper einher, was zu einer Reduktion der Nährstoffversorgung der Bandscheibe und damit zu einem Untergang der matrixbildenden Zellen und konsekutiv zu einem Flüssigkeitsverlust in der Bandscheibe führt. Als Folge nimmt die Belastung der Bandscheibe weiter ab. Die aktuellen Therapieoptionen umfassen sowohl die konservative als auch die operative Therapie, wobei allerdings nur die Folgen der Degeneration behandelt werden. Ziel einer Zelltransplantation ist es, der Bandscheibe wieder matrixbildende Zellen zur Verfügung zu stellen, damit die nutritiven Veränderungen auszugleichen und eventuell reversibel zu machen. Dieser Effekt konnte im Tierversuch nachgewiesen werden, woraufhin eine klinische Studie initiiert wurde. Im Rahmen der publizierten klinischen prospektiven, randomisierten Studie konnte gezeigt werden, dass die Transplantation autologer Chondrozyten, die bei einer notwendigen operativen Therapie eines sequestrierten Bandscheibenvorfalls gewonnen wurden, einen sowohl klinisch als auch bildmorphologisch positiven Effekt auf die degenerierten Bandscheiben hat. Es kam zu einer signifikanten Reduktion der Schmerzsymptomatik und einer Steigerung der Lebensqualität in der Gruppe der transplantierten Patienten. Die Bandscheibenhöhe zeigt sich stabil über den Beobachtungszeitraum von 2 Jahren.

The research of new therapeutic strategies for cardiovascular diseases has seen in the last 15 years the introduction of a new participant: pericardial adipose tissue (PAT). This tissue is able to modulate cardiac function and its volume was sometimes linked to risk of cardiovascular diseases. Moreover, adipose-derived stem cells (ASCs) isolated from PAT are considered as the best suitable for new regenerative strategies aiming at healing ischemic myocardium. Although the interests in understanding the functions and the formation of pericardial adipose tissue are high, the current knowledge on this tissue is still scarce. In this work, the starting point was the consideration that nucleotide receptors are established regulators of many biological functions, including the differentiation of adult mesenchymal stem cells, immunity and inflammatory process. The P2Y4 receptor has been recently recognized as a negative regulator of cardiac fat formation and ASCs differentiation. The purpose of this thesis was to analyze the involvement of the nucleotide receptor P2Y2 in the formation of pericardial adipose tissue (PAT) and its ASCs differentiation. We also investigated the possible contribution of this receptor to the functions of recently discovered fat-associated lymphoid clusters (FALCs). Our study analyzed the PAT of mice deficient for P2Y2 at basal conditions and in a model of myocardial infarction. P2Y2-null mice showed a lower mass of PAT compared to WT, which was correlated with decreased adipogenic differentiation and maturation potential of pericardial ASCs in vitro. PAT of basal P2Y2-deficient mice displayed a reduced density of FALCs due to a reduced number of B cells. RNA-sequencing experiments identified many P2Y2 target genes in PAT linked to immunomodulation. We identified a polarization of FALCs macrophages towards anti-inflammatory M2c subtype in P2Y2-null mice. We correlated it with a decreased number of follicular helper T cells, known to contribute to B cell expansion in germinal centers. These data could be correlated with increased apoptosis of B lymphocytes. The data obtained using the mouse infarct model confirmed an expected enlargement of pericardial FALCs in ischemic conditions. P2Y2-null mice were characterized by a reduced expansion of B cells and myeloid cells migration in PAT. These results suggested a participation of P2Y2 receptor in regulating the post-MI inflammatory response by modulating the leukocytes populations in the pericardial adipose tissue’s lymphoid clusters. The effect of P2Y2 on PAT post-ischemic inflammatory state could contribute to the P2Y2-mediated cardioprotective effect of UTP described in previous literature. Our study defines P2Y2 nucleotide receptor as a regulator of the formation of pericardial fat and its inflammatory status in ischemic conditions. P2Y2 receptor could represent an interesting therapeutic target for the regulation of PAT functions before and after MI. In general, a better comprehension of PAT and its consideration in the post-ischemic regeneration process could lead to the development of new therapeutic strategies for treating cardiovascular diseases and the adjustment of existing therapies. / Durant les 15 dernières années, un nouvel arrivant a fait son apparition dans la recherche de nouvelles approches thérapeutiques dans le domaine cardiovasculaire: le tissu adipeux cardiaque. Ce tissu est capable de moduler les fonctions cardiaques et son volume a pu être associé parfois à un risque de maladie cardiovasculaire. De plus, les cellules souches dérivées du tissu adipeux (ASCs) cardiaque sont considérées comme les mieux appropriées pour des stratégies thérapeutiques visant la réparation du myocarde ischémié. Bien que la compréhension de la fonction et de la formation du tissu adipeux cardiaque présente un intérêt majeur, la connaissance actuelle de ce tissu particulier est encore assez limitée. Pour le présent travail, le point de départ a été l’observation que les récepteurs nucléotidiques sont des régulateurs établis de nombreuses fonctions biologiques, incluant la différentiation des cellules souches mésenchymateuses et plus généralement la régulation de la réponse immune et inflammatoire. Le récepteur P2Y4 a été récemment reconnu comme un régulateur négatif de la formation du tissu adipeux cardiaque et de la différentiation des ASCs. Le but de cette thèse a été l’étude de l’implication du récepteur nucléotidiques P2Y2 dans la formation du tissu adipeux péricardique (TAP) et la différentiation des ASCs. Nous avons également investigué la contribution possible de ce récepteur dans la fonction des structures leucocytaires associées au tissu adipeux appelées FALCS pour fat-associated lymphoid clusters.Nous avons étudié le TAP de souris déficientes pour le récepteur P2Y2 à l’état de base et dans un modèle d’infarctus du myocarde. Les souris P2Y2 knock-out (KO) présentent une masse réduite du TAP corrélée avec le fait que l’absence du P2Y2 diminue la différentiation adipogénique et le potentiel de maturation des ASCs péricardiques in vitro. Le PAT des souris P2Y2 KO présentent une diminution de la densité de FALCs à l’état de base, principalement due à un nombre réduit de lymphocytes B, potentiellement corrélé à une apoptose accrue observée dans ces cellules. Nos expériences de RNA-sequencing ont identifié de nombreux gènes cibles du P2Y2 dans le PAT impliqués dans l’immunomodulation. Nous avons identifié une polarisation des macrophages de type M2c dans les FALCs de souris P2Y2 KO. Nous l’avons corrélée avec une diminution des lymphocytes T helper folliculaires connus pour contribuer à l’expansion des lymphocytes B dans les centres germinaux. Les données obtenues dans le modèle d’infarctus chez la souris ont confirmé une augmentation des FALCs péricardiques dans les conditions d’ischémie cardiaque. Les souris P2Y2 KO sont caractérisées par une expansion réduite des lymphocytes B et des cellules myéloïdes dans le TAP. Ces résultats suggèrent une participation du récepteur P2Y2 dans la régulation de la réponse inflammatoire post-infarctus par la modulation des populations leucocytaires dans les clusters lymphocytaires du tissu adipeux cardiaque. L’effet du P2Y2 sur l’état inflammatoire post-ischémique pourrait contribuer à l’effet cardioprotecteur de l’UTP médié par le P2Y2 et précédemment décrit dans la littérature.Notre étude définit le récepteur nucléotidique P2Y2 comme un régulateur de la formation du tissu adipeux péricardique et de son niveau inflammatoire dans des conditions ischémiques. Le récepteur P2Y2 pourrait représenter une cible thérapeutique intéressante pour la régulation des fonctions du PAT avant et après infarctus du myocarde. Plus généralement, une meilleure compréhension du tissu adipeux cardiaque et de son implication dans le processus de régénération cardiaque pourrait mener au développement de nouvelles stratégies thérapeutiques dans le traitement de maladies cardiovasculaires et à l’ajustement de thérapies déjà existantes. / Doctorat en Sciences biomédicales et pharmaceutiques (Médecine) / info:eu-repo/semantics/nonPublished

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PhD (Department of Biotechnology, Faculty of Applied and Computer Sciences), Vaal University of Technology. / Tulbaghia violacea Harv. has found extensive uses in traditional medicine for the treatment of numerous ailments among which are tuberculosis, oesophageal cancer, diabetes and cardiovascular diseases. Current reports show that cardiovascular diseases are now the primary cause of mortality worldwide. Thus, the potential of T. violacea plant extracts against cardiovascular diseases should be explored. The objectives of this study were, (i) to conduct qualitative and quantitative preliminary phytochemical screening of T. violacea aqueous leaf extracts, (ii) to conduct Gas chromatography–mass spectrometry (GC-MS) analysis for screening of compounds present in the plant extract, (iii) to evaluate the antioxidant activity of the T. violacea crude extracts using the DPPH:1.1-diphenyl-2-picrylhydrazyl and ABTS: 2,2-azino-bis 3-ethylebenzthiazoline-6-sulfonic acid assays, (iv) to evaluate the antimicrobial activity of the T. violacea crude extracts using disk diffusion and Minimum inhibitory concentration/Minimum bactericidal concentration (MIC/MBC), (v) to evaluate the antithrombogenic properties of T. violacea crude extracts on polystyrene, (vi) to fabricate polycaprolactone (PCL) and PCL-T. violacea incorporated scaffolds, (vii) to evaluate the antithrombogenic properties of T. violacea crude extracts on the fabricated PCL and PCL-T. violacea fabricated scaffolds and, (viii) to evaluate the growth and differentiation of adipose derived stem cells (ADSCs) on the fabricated scaffolds. The qualitative and quantitative phytochemical screening was conducted using standard procedures. Folin-Ciocalteu method was used to evaluate both total phenolic content (TPC) and total tannin content (TTC), the Aluminium chloride method was used for total flavonoid content (TFC) and GC-MS was used to screen for compounds present in the plant extract. The antioxidant activity was evaluated using DPPH and ABTS and the antimicrobial activity was evaluated using disc diffusion and MIC/MBC assays. The antithrombogenic properties of the T. violacea aqueous leaf extracts was then evaluated using platelet activation and whole blood clotting kinetics on polystyrene discs which have been reported to induce platelet activation. The experiment was performed in the absence and presence of 100 and 1000 μg/ml T. violacea plant extracts for both the platelet activation study which used blood plasma and the whole blood clotting kinetics assay which used fresh whole blood. Platelet adhesion was evaluated using fluorescence microscopy and a scanning electron microscope (SEM) was used to evaluate their morphology. Three scaffolds designated as PCL, 10% Tvio and 15% Tvio were fabricated which consisted of a 10% PCL powder and 10% as well as 15% T. violacea aqueous plant extract with respect to the PCL powder weight. The scaffolds were then characterized using Fourier-transform infrared spectroscopy (FTIR) and Energy-dispersive x-ray spectroscopy (EDS). The scaffolds were then evaluated for their antithrombogenic properties in the presence and absence of 100 and 1000 μg/ml T. violacea plant extracts. Platelet adhesion was evaluated using a fluorescent microscope and the morphology was evaluated using SEM. For the cell study, adipose derived stem cells (ADSCs) were cultured on the designed scaffolds and evaluated for their toxicity, viability, adhesion, proliferation, morphology and differentiation into osteoblasts over a period of 3 weeks. Lactate dehydrogenase (LDH) assay was used for toxicity studies, alamar blue assay was used for viability, fluorescence microscopy was used to evaluate cellular adhesion and proliferation while the alkaline phosphate (ALP) assay was used to evaluate differentiation of the cells into osteoblasts. Cell morphology was evaluated using SEM. Phytochemical screening of the prepared T. violacea aqueous extract revealed the presence of terpenoids, flavonoids, cardiac glycosides, saponins, protein, phenols, tannins, carbohydrates and amino acids. This is the first study that has identified the presence of carbohydrates and amino acids in T. violacea aqueous leaf extracts. Different concentrations of 0.1, 1.0 and 10 mg/ml of plant extract were used to conduct the quantitative phytochemical screening assays. There was a concentration dependent increase in the amount of phenols, tannins and flavonoids as the concentration of the plant extracts increased. This was the first study that evaluated the total tannic content of T. violacea plant extracts. The amount of total phenols was higher than that of flavonoids and tannins at every concentration range studied followed by the total flavonoids and lastly total tannins. The GC-MS analysis showed the presence of 33 compounds among which were 2,4 – Dithiapentate - 2,2-dioxide, Cannabidiol, 2,4,5,7 –Tetrathiaoctane and 2,4,5,7 - Tetrathiaoctane 2-dioxide. The presence of sulphur compounds support the characteristic garlic-like smell as well as some of the biological activities of T. violacea plant extracts. The antioxidant activities based on DPPH (0.49 mg/ml) and ABTS (0.24 mg/ml) suggest that T. violacea can be used as potential antioxidant agents. For the antimicrobial activity using disc diffusion, the extracts exhibited appreciable antibacterial activities against Bacillus subtilis, Serratia marcescens, Staphylococcus aureus and S. epidermidis. The highest zone of inhibition was observed for S. epidermidis at 19.50 ± 0.87 mm. The MIC results revealed that the plant extract of T. violacea was moderately active against B. subtilis, S. aureus, S. epidermidis, E. coli, and S. marcescens with MIC value of 2.5 mg/ml. However, the antimicrobial effect of the extract on S. epidermidis was bactericidal when compared to the bacteriostatic effect on the other active microorganisms. The antithrombogenic results on the polystyrene discs showed a significant reduction in the number of platelets that adhered on the polystyrene surfaces treated with plasma mixed with 100 μg/ml of plant extract when compared to the untreated control and the 1000 μg/ml treatment. For the 1000 μg/ml treatment, there was a significant increase in the number of platelets that adhered to polystyrene surfaces. These results were confirmed by the fluorescence and SEM results which showed a higher platelet count for the 1000 μg/ml treatment when compared to the other groups. The whole blood clotting kinetics study showed delayed blood clotting with the 100 μg/ml treatment over a period of 60 min when compared to the untreated control and the 1000 μg/ml treatment. These results correspond with the lower platelet adhesion observation and thus confirm the anticlotting properties of T. violacea aqueous leaf extracts at lower concentrations. The mean diameter of the scaffolds was recorded on the SEM as 275.60 ± 60.65 nm, 193 ± 30 nm and 537 ± 138 nm for the PCL, 10% Tvio and 15% Tvio scaffolds, respectively. The FTIR spectrum revealed the presence of amide groups as well hydroxyl O–H stretching groups which were the characteristic groups for the presence of T. violacea plant extracts in the polycaprolactone. The EDS results showed the presence of potassium, chlorine and sulphur compounds which were only present in the T. violacea scaffolds in addition to the carbon, oxygen and silicon observed in the PCL scaffold. The fabricated scaffolds were then used to evaluate platelet adhesion and activation on blood plasma in the absence and presence of 100 and 1000 μg/ml T. violacea aqueous leaf extracts. The results showed that the 10% Tvio scaffold was more effective in inhibiting platelet adhesion and activation at every treatment group especially when plasma was used in the absence of T. violacea plant extracts. A similar observation to the polystyrene study was observed were addition of 1000 μg/ml of plant extract resulted in the highest number of activated platelets. The study suggests the potential of the 10% Tvio scaffold in the prevention of platelet adhesion and aggregation. The in vitro cell adhesion, proliferation and differentiation of adipose derived stem cells (ADSCs) on the fabricated T. violacea loaded PCL nanofibers was then evaluated. The LDH assay illustrated less activity on the 10% Tvio scaffold when compared to PCL and 15% Tvio scaffolds however, none of the scaffolds were considered as toxic. The alamar blue assay was used for viability after 4 and 7 days of culture. The results showed a significant increase in cell viability for all scaffolds from day 4 to day 7 with the 10% Tvio scaffold having the highest overall cell viability for both day 4 and day 7 of cell cultures. Immunofluorescence staining was then used to count the number of cells using DAPI (4′,6-diamidino-2-phenylindole) stained images and illustrated that the T. violacea incorporated scaffolds supported better cell growth compared to the PCL scaffold. Cell morphology on the T. violacea scaffolds was denser and spread out into cellular extensions when compared to the PCL scaffold after 7 days of cell culture, supporting the higher number of adhered cells from the fluorescence results. For the long term cell study after week 1 and 3, the ALP results showed a significant difference in ALP activity between week 1 and week 3 for all scaffolds. The highest ALP activity was observed for the 15% Tvio scaffolds which is a marker for initial phase of bone matrix deposition. The designed T. violacea scaffolds supported better cell growth compared to the PCL scaffold and their morphology was more spread out and covered the entire surface of the scaffolds after week 3. Lastly, the cell count and osteocalcin differentiation was more prominent on 10% Tvio scaffold indicating higher levels of the protein marker for bone formation. Thus, supporting the use of the 10% Tvio scaffold for long-term cell studies. In conclusion, the results of this study indicated that the aqueous extract of T. violacea is rich is phytochemicals and also possess a broad range of pharmaceutically important compounds which may be attributed to the high antioxidant and antimicrobial activities identified. The results from this study suggest that T. violacea aqueous extracts have antithrombogenic properties at lower concentrations. Scaffolds fabricated with the incorporation of T. violacea plant extract also confirm the potential antiplatelet activity of the fabricated 10% Tvio scaffold. The results also suggest the potential of the fabricated 10% Tvio scaffold to enhance cell adhesion, proliferation and differentiation over long-term cell studies. It can thus be recommended that T. violacea may be useful for tissue engineering applications and bone repair with prospects of preventing cardiovascular diseases associated with bone defects. This research study has provided the foundation for clinical evaluation and outlined the potential effects of T. violacea aqueous leaf extracts as a clinical drug.