RESUMO
OBJECTIVE: This study aims to develop a compound biomaterial to achieve effective soft tissue regeneration. METHODOLOGY: Compound hyaluronic acid (CHA) and liquid horizontal-platelet-rich fibrin (H-PRF) were mixed at a ratio of 1:1 to form a CHA-PRF gel. Human gingival fibroblasts (HGFs) were used in this study. The effect of CHA, H-PRF, and the CHA-PRF gel on cell viability was evaluated by CCK-8 assays. Then, the effect of CHA, H-PRF, and the CHA-PRF gel on collagen formation and deposition was evaluated by qRTâPCR and immunofluorescence analysis. Finally, qRTâPCR, immunofluorescence analysis, Transwell assays, and scratch wound-healing assays were performed to determine how CHA, H-PRF, and the CHA-PRF gel affect the migration of HGFs. RESULTS: The combination of CHA and H-PRF shortened the coagulation time of liquid H-PRF. Compared to the pure CHA and H-PRF group, the CHA-PRF group exhibited the highest cell proliferation at all time points, as shown by the CCK-8 assay. Col1a and FAK were expressed at the highest levels in the CHA-PRF group, as shown by qRTâPCR. CHA and PRF could stimulate collagen formation and HGF migration, as observed by fluorescence microscopy analysis of COL1 and F-actin and Transwell and scratch healing assays. CONCLUSION: The CHA-PRF group exhibited greater potential to promote soft tissue regeneration by inducing cell proliferation, collagen synthesis, and migration in HGFs than the pure CHA or H-PRF group. CHA-PRF can serve as a great candidate for use alone or in combination with autografts in periodontal or peri-implant soft tissue regeneration.
Assuntos
Movimento Celular , Proliferação de Células , Sobrevivência Celular , Fibroblastos , Gengiva , Ácido Hialurônico , Fibrina Rica em Plaquetas , Regeneração , Ácido Hialurônico/farmacologia , Humanos , Fibroblastos/efeitos dos fármacos , Gengiva/efeitos dos fármacos , Gengiva/citologia , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Regeneração/efeitos dos fármacos , Fatores de Tempo , Movimento Celular/efeitos dos fármacos , Reprodutibilidade dos Testes , Imunofluorescência , Reação em Cadeia da Polimerase em Tempo Real , Colágeno , Teste de Materiais , Cicatrização/efeitos dos fármacos , Materiais Biocompatíveis/farmacologia , Colágeno Tipo I/análiseAssuntos
Colágeno Tipo I , Peptídeos , Colágeno Tipo I/sangue , Colágeno Tipo I/análise , Humanos , Peptídeos/análise , Peptídeos/química , Automação , IsomerismoRESUMO
Adult tendons are highly differentiated. In mature individuals, tendon healing after an injury occurs through fibrotic tissue formation. Understanding the intrinsic reparative properties of fetal tendons would help to understand the maturation tissue process and tendon tissue repair. The present study evaluated the evolution of histoarchitecture, cellularity and the distribution of collagens I, III and V in the posterior tibial tendon in human fetuses at different gestational ages. Morphological profiles were assessed in nine fresh spontaneously aborted fetuses (Group I: five fetuses aged between 22 and 28 weeks of gestation; Group II: four fetuses aged between 32 and 38 weeks of gestation), characterized by a combination of histology, fluorescence and immunohistochemistry. In Group I, the posterior tibial tendon showed statistically significant greater cellularity and presence of collagen III and V than in Group II tendon, which showed a predominance of collagenous I and a better organization of the extracellular matrix compared with Group I tendons. In addition, a statistically significant higher rate of CD90, a marker of mesenchymal cells, was found in Group I tendons. In fetuses with gestational age between 22 and 28 weeks, the posterior tibialis tendons showed a thin and disorganized fibrillar structure, with an increase in collagen III and V fibers and mesenchymal cells. In the posterior tibialis tendons of fetuses with gestational age between 32 and 38 weeks, the fibrillar structure was thicker with a statistically significant increase in type I collagen and decreased cellularity.
Assuntos
Colágeno Tipo I , Tendões , Adulto , Humanos , Lactente , Colágeno Tipo I/análise , Tendões/patologia , Matriz Extracelular/química , Colágeno/química , FetoRESUMO
SUMMARY: Severe muscle injuries are common in accidents and have a delayed recovery of muscle integrity. In these cases, muscle suture surgery is the standard treatment. However, Platelet Rich Plasma (PRP), has been widely used in orthopedic injuries due to its growth factors. Thus, the objective of the study will be to analyze the association of suture and PRP techniques in the collagen and tenacity of the injured muscle. Were used seventy rats, divided into five groups: control (C), injury control (CI), injury and suture (IS), injury and PRP (IP), injury, suture, and PRP (ISP). Were sectioned approximately 50 % of the width and 100 % of the thickness of the gastrocnemius muscle. The homologous PRP was applied 24h after the injury. On the 7th day after the injury, the animals were euthanized and their muscles subjected to mechanical testing to measure tenacity or collagen analysis to calculate the ratio between type I and III collagen. The results show a significant decrease (p <0.05) in the values of the relationship between collagens in all injured groups (CI, IS, IP, ISP) compared to group C. In injured groups, the tenacity was significantly (p <0.05) reduced compared to the control group, with no observed difference between treatments and injured groups. The amount of collagen in the injured area has increased, but it did not affect the tenacity of the muscles, which was reduced.
RESUMEN: Las lesiones musculares graves son comunes durante los accidentes y la integridad del músculo está sujeta a una larga recuperación. En esos casos la cirugía, para la sutura del músculo, es el tratamiento común, no obstante el plasma rico en plaquetas (PRP) ha sido utilizado recientemente en lesiones ortopédicas, debido a sus factores del crecimiento. El objetivo del estudio fue analizar la asociación de las técnicas de sutura y PRP en la histología y tenacidad de músculo lesionado. Fueron utilizadas 70 ratas distribuidas en cinco grupos: control (C), control lesión (CL), lesión y sutura (LS), lesión y PRP (LPRP), lesión, sutura y PRP (LSPRP). Aproximadamente en la lesión, el 50 % de la longitud y el 100 % del espesor del músculo gastrocnemio fueron seccionados. El PRP homólogo fue aplicado 24 horas después de la lesión. En el 7º día después de la lesión los animales fueron eutanasiados y las muestras fueran sometidas al ensayo mecánico para la medición de la tenacidad y análisis del colágeno, para realizar el cálculo de la relación entre los colágenos I y III. Los resultados demostraron una reducción significativa (p<0,05) en los valores de la relación entre los colágenos en todos los grupos lesionados en relación al grupo C. La tenacidad fue (p<0,05) reducida significativamente en los grupos lesionados en relación al grupo control, sin diferencia entre los tratados. En la lesión muscular hubo disminución de los valores de colágeno, aunque en los tratamientos se observó elevación de la cantidad de colágeno en la área lesionada, esta no tuvo efecto en la tenacidad de los músculos que fue disminuida en la lesión.
Assuntos
Animais , Masculino , Ratos , Colágeno/análise , Músculo Esquelético/lesões , Plasma Rico em Plaquetas , Doenças Musculares/terapia , Suturas , Ratos Wistar , Lesões dos Tecidos Moles/terapia , Colágeno Tipo I/análise , Colágeno Tipo III/análiseRESUMO
PURPOSE: To evaluate the in vivo response of photobiomodulation therapy associated with norbixin-based poly(hydroxybutyrate) membrane (PHB) in tenotomized calcaneal tendon. METHODS: Thirty rats were randomly allocated to six groups (n=5 each): LED groups (L1, L2 and L3) and membrane + LED groups (ML1, ML2 and ML3). The right calcaneal tendons of all animals were sectioned transversely and were irradiated with LED daily, one hour after surgery every 24 hours, until the day of euthanasia. At the end of the experiments the tendons were removed for histological analysis. RESULTS: The histological analysis showed a significant reduction in inflammatory cells in the ML1, ML2 and ML3 groups (p=0.0056, p=0.0018 and p<0.0001, respectively) compared to those in the LED group. There was greater proliferation of fibroblasts in the ML1 (p<0.0001) and L3 (p<0.0001) groups. A higher concentration of type I collagen was also observed in the ML1 group (p=0.0043) replacing type III collagen. CONCLUSION: Photobiomodulation in association with norbixin-based PHB membrane led to control of the inflammatory process. However, it did not favor fibroblast proliferation and did not optimize type I collagen formation in the expected stage of the repair process.
Assuntos
Tendão do Calcâneo/efeitos da radiação , Carotenoides/farmacologia , Hidroxibutiratos/farmacologia , Terapia com Luz de Baixa Intensidade/métodos , Tendinopatia/radioterapia , Tenotomia/métodos , Tendão do Calcâneo/efeitos dos fármacos , Tendão do Calcâneo/cirurgia , Animais , Colágeno/farmacologia , Colágeno Tipo I/análise , Colágeno Tipo I/efeitos dos fármacos , Colágeno Tipo III/análise , Colágeno Tipo III/efeitos dos fármacos , Avaliação Pré-Clínica de Medicamentos , Fibroblastos/química , Fibroblastos/efeitos dos fármacos , Masculino , Proibitinas , Distribuição Aleatória , Ratos , Ratos Wistar , Cicatrização/efeitos dos fármacos , Cicatrização/efeitos da radiaçãoRESUMO
OBJECTIVE: Previous studies have reported a close relationship between malignant mesothelioma (MM) and the immune matricial microenvironment (IMM). One of the major problems in these studies is the lack of adequate adjustment for potential confounders. Therefore, the aim of this study was to identify and quantify risk factors such as IMM and various tumor characteristics and their association with the subtype of MM and survival. METHODS: We examined IMM and other tumor markers in tumor tissues from 82 patients with MM. These markers were evaluated by histochemistry, immunohistochemistry, immunofluorescence, and morphometry. Logistic regression analysis, cluster analysis, and Cox regression analysis were performed. RESULTS: Hierarchical cluster analysis revealed two clusters of MM that were independent of clinicopathologic features. The high-risk cluster included MM with high tumor cellularity, high type V collagen (Col V) fiber density, and low CD8+ T lymphocyte density in the IMM. Our results showed that the risk of death was increased for patients with MM with high tumor cellularity (OR = 1.63, 95% CI = 1.29-2.89, P = .02), overexpression of Col V (OR = 2.60, 95% CI = 0.98-6.84, P = .04), and decreased CD8 T lymphocytes (OR = 1.001, 95% CI = 0.995-1.007, P = .008). The hazard ratio for the high-risk cluster was 2.19 (95% CI = 0.54-3.03, P < .01) for mortality from MM at 40 months. CONCLUSION: Morphometric analysis of Col V, CD8+ T lymphocytes, and tumor cellularity can be used to identify patients with high risk of death from MM.
Assuntos
Biomarcadores Tumorais/análise , Mesotelioma Maligno/mortalidade , Microambiente Tumoral , Linfócitos T CD8-Positivos , Colágeno Tipo I/análise , Colágeno Tipo V/análise , Colágeno Tipo V/metabolismo , Feminino , Imunofluorescência , Humanos , Imuno-Histoquímica , Hibridização In Situ , Contagem de Linfócitos , Masculino , Mesotelioma Maligno/imunologia , Mesotelioma Maligno/metabolismo , Mesotelioma Maligno/patologia , Pessoa de Meia-Idade , Análise de Regressão , Estudos Retrospectivos , Medição de Risco , Fatores de Risco , Análise Serial de Tecidos , Microambiente Tumoral/imunologiaRESUMO
Although dental implants and bone regenerative procedures are important approaches for the reestablishment of esthetics and function in young patients with a history of generalized aggressive periodontitis (GAP), no predictable outcomes have been reported, and the host osteo-immunoinflammatory response may play a relevant role in this context. In view of the lack of molecular investigations into the bone tissue condition of young patients with periodontitis, the aim of this study was to evaluate the gene expression of bone-related factors in this population. Bone biopsies were obtained from the posterior mandible in 16 individuals previously diagnosed with GAP and on periodontal support therapy and from 17 periodontally healthy (PH) patients. The gene expression of tumor necrosis factor (TNF)-α, transforming growth factor (TGF)-ß, receptor activator of the NF-κB ligand (RANKL), osteoprotegerin (OPG), osteocalcin (OC), bone sialoprotein (BSP), and type I collagen (COL-I), important biomarkers of bone turnover, was evaluated by qRT-PCR. Lower TGF-ß and OPG mRNA levels were observed in GAP patients compared to PH individuals (p ≤ 0.05). There were no between-group differences in levels of TNF-α, BSP, RANKL, OC, or COL-I mRNA (p>0.05). In young adults, a history of periodontal disease can negatively modulate the gene expression of important bone-related factors in alveolar bone tissue. These molecular outcomes may contribute to the future development of therapeutic approaches to benefit bone healing in young patients with history of periodontitis via modulation of osteo-immuno-inflammatory biomarkers.
Assuntos
Periodontite Agressiva/genética , Expressão Gênica , Adulto , Periodontite Agressiva/metabolismo , Processo Alveolar/química , Biomarcadores , Colágeno Tipo I/análise , Colágeno Tipo I/genética , Estudos Transversais , Feminino , Humanos , Sialoproteína de Ligação à Integrina/análise , Sialoproteína de Ligação à Integrina/genética , Masculino , Osteocalcina/análise , Osteocalcina/genética , Osteoprotegerina/análise , Osteoprotegerina/genética , Ligante RANK/análise , Ligante RANK/genética , Reação em Cadeia da Polimerase em Tempo Real , Valores de Referência , Método Simples-Cego , Estatísticas não Paramétricas , Fator de Crescimento Transformador beta/análise , Fator de Crescimento Transformador beta/genética , Fator de Necrose Tumoral alfa/análise , Fator de Necrose Tumoral alfa/genética , Adulto JovemRESUMO
PURPOSE: To determine the efficacy of norbixin-based poly(hydroxybutyrate) (PHB) membranes for Achilles tendon repair. METHODS: Thirty rats were submitted to total tenotomy surgery of the right Achilles tendon and divided into two groups (control and membrane; n = 15 each), which were further subdivided into three subgroups (days 7, 14, and 21; n = 5 each). Samples were analyzed histologically. RESULTS: Histological analysis showed a significant reduction in inflammatory infiltrates on days 7, 14 (p < 0.0001 for both), and 21 (p = 0.0004) in the membrane group compared to that in the control group. There was also a significant decrease in the number of fibroblasts in the control group on days 7, 14 (p < 0.0001), and 21 (p = 0.0032). Further, an increase in type I collagen deposition was observed in the membrane group compared to that in the control group on days 7 (p = 0.0133) and 14 (p = 0.0107). CONCLUSION: Treatment with norbixin-based PHB membranes reduces the inflammatory response, increases fibroblast proliferation, and improves collagen production in the tendon repair region, especially between days 7 and 14.
Assuntos
Tendão do Calcâneo/efeitos dos fármacos , Tendão do Calcâneo/cirurgia , Carotenoides/farmacologia , Hidroxibutiratos/farmacologia , Poliésteres/farmacologia , Tenotomia/métodos , Tendão do Calcâneo/patologia , Animais , Colágeno Tipo I/análise , Colágeno Tipo I/efeitos dos fármacos , Colágeno Tipo III/análise , Colágeno Tipo III/efeitos dos fármacos , Fibroblastos/efeitos dos fármacos , Humanos , Masculino , Proibitinas , Ratos Wistar , Valores de Referência , Regeneração/efeitos dos fármacos , Reprodutibilidade dos Testes , Fatores de Tempo , Resultado do TratamentoRESUMO
Objective: The objective of this study was to evaluate the action of soy isoflavones (ISO) and 17ß-estradiol on collagen I (CollI) and sulfated glycosaminoglycans (GAGs) in the bone matrix of diabetic rats.Methods: Sixty adult female rats (Rattus norvegicus albinus) underwent ovariectomy, and then were randomized into six groups of 10 animals each: GI, sham control ovariectomized animals; GII, sham control diabetic (DM) ovariectomized animals; GIII, control ovariectomized animals receiving propylene glycol vehicle; GIV, control ovariectomized DM animals receiving propylene glycol vehicle; GV, ovariectomized DM animals treated with ISO (150 mg/kg by gavage); and GVI, ovariectomized DM animals treated with estrogen (17ß-estradiol, 10 mg/kg, subcutaneously). 17ß-Estradiol was used as a positive control when compared with ISO. To obtain significant depletion of the estrogen levels and subsequent bone loss, a postsurgical period of 90 days was observed. Treatments occurred during 30 consecutive days. After euthanasia, shafts of the animals' femurs were immersed in liquid nitrogen for molecular biology analysis, and the distal femurs were removed and processed for paraffin embedding.Results: ISO (GV) and 17ß-estradiol (GVI) improved bone formation, increasing GAGs and CollI formation when compared to the control group (GIV) (p < 0.05).Conclusions: ISO and 17ß-estradiol contribute to the decrease of bone loss in diabetic rats.
Assuntos
Osso e Ossos/metabolismo , Estradiol/farmacologia , Estrogênios/farmacologia , Isoflavonas/química , Animais , Colágeno Tipo I/análise , Diabetes Mellitus Experimental , Diabetes Mellitus Tipo 1/metabolismo , Estradiol/metabolismo , Estrogênios/metabolismo , Feminino , Glicosaminoglicanos/análise , Humanos , Isoflavonas/metabolismo , Ovariectomia , Pós-Menopausa , Distribuição Aleatória , RatosRESUMO
Abstract Although dental implants and bone regenerative procedures are important approaches for the reestablishment of esthetics and function in young patients with a history of generalized aggressive periodontitis (GAP), no predictable outcomes have been reported, and the host osteo-immunoinflammatory response may play a relevant role in this context. In view of the lack of molecular investigations into the bone tissue condition of young patients with periodontitis, the aim of this study was to evaluate the gene expression of bone-related factors in this population. Bone biopsies were obtained from the posterior mandible in 16 individuals previously diagnosed with GAP and on periodontal support therapy and from 17 periodontally healthy (PH) patients. The gene expression of tumor necrosis factor (TNF)-α, transforming growth factor (TGF)-β, receptor activator of the NF-κB ligand (RANKL), osteoprotegerin (OPG), osteocalcin (OC), bone sialoprotein (BSP), and type I collagen (COL-I), important biomarkers of bone turnover, was evaluated by qRT-PCR. Lower TGF-β and OPG mRNA levels were observed in GAP patients compared to PH individuals (p ≤ 0.05). There were no between-group differences in levels of TNF-α, BSP, RANKL, OC, or COL-I mRNA (p>0.05). In young adults, a history of periodontal disease can negatively modulate the gene expression of important bone-related factors in alveolar bone tissue. These molecular outcomes may contribute to the future development of therapeutic approaches to benefit bone healing in young patients with history of periodontitis via modulation of osteo-immuno-inflammatory biomarkers.