The enteric glia: identity and functions.

Enteric glial cells were first described at the end of the 19th century, but they attracted more interest from researchers only in the last decades of the 20th. Although, they have a different embryological origin, the enteric GLIA share many characteristics with astrocytes, the main glial cell type of the central nervous system (CNS), such as in their expression of the same markers and in their functions. Here we review the construction of the enteric nervous system (ENS), with a focus on enteric glia, and also the main studies that have revealed the action of enteric glia in different aspects of gastrointestinal tract homeostasis, such as in the intestinal barrier, in communications with neurons, and in their action as progenitor cells. We also discuss recent discoveries about the roles of enteric glia in different disorders that affect the ENS, such as degenerative pathologies including Parkinson's and prion diseases, and in cases of intestinal diseases and injury.

Approaches to preserve human osteochondral allografts.

Osteochondral defects may progress to osteoarthritis. Many attempts have been developed to overcome this issue, including osteochondral autografts and allografts. The goal of this study was to develop a new protocol for storage of human osteochondral allografts. Osteochondral plugs were randomly allocated in the following groups: control, immediate freezing up to -70 °C, cooling at 4 °C, and storage at 37 °C. Samples from the cooling at 4 °C and storage at 37 °C groups were stored in tubes containing medium plus human albumin and analyzed after 1, 3, and 14 days. The frozen groups' samples were cryopreserved for 1 year in cryotubes containing medium only (FM), medium plus human albumin (FA), and medium plus human albumin and glucose (FG) and were then analyzed. Analysis involved histological study with hematoxylin-eosin and Safranin O and a modified Live/Dead assay. In samples stored both at 37 and 4 °C, analysis showed statistically significant higher cellular mortality at 14 days compared to 1 and 3 days, but mortality in the 4 °C group was lower. In the freezing protocols, the FA group showed less cellular mortality than the FM and FG groups. Cooling at 4 °C offers better preservation capacity than storage at 37 °C, but both offer the capacity for preservation for 14 days. Adding human albumin to the storage medium is useful in reducing cellular mortality in samples frozen for 1 year.

New strategy to control cell migration and metastasis regulated by CCN2/CTGF.

Connective tissue growth factor (CTGF)/CCN family member 2 (CCN2) is a CCN family member of matricellular signaling modulators. It has been shown that CCN2/CTGF mediates cell adhesion, aggregation and migration in a large variety of cell types, including vascular endothelial cells, fibroblasts, epithelial cells, aortic smooth muscle and also pluripotent stem cells. Others matricellular proteins are capable of interacting with CCN2/CTGF to mediate its function. Cell migration is a key feature for tumor cell invasion and metastasis. CCN2/CTGF seems to be a prognostic marker for cancer. In addition, here we intend to discuss recent discoveries and a new strategy to develop therapies against CCN2/CTGF, in order to treat cancer metastasis.

Conservative Treatment of Camptodactyly with use of Orthoses: A Retrospective Cohort.

Objective To evaluate the outcomes of conservative treatment using static orthoses manufactured by the Occupational Therapy Sector of our institution in participants with camptodactyly types I, II, and III in their rigid or flexible forms, to describe the demographic and clinical data, and to analyze the number of dropouts during the treatment period. Methods The Ethics in Human Research Committee approved the project under protocol number CAAE 20300419.3.0000.5273. All medical records used in the research were made available by our institution. In the present retrospective study, we did not use the informed consent form due to the impossibility of contacting the high number of participants. The study included medical records of 38 participants treated at the Occupational Therapy Outpatient Clinic from 2013 to 2019. Results Of the 54 fingers treated with orthoses, 38 were completely corrected. The rates of correction were as follows: type I in its rigid form - 100% type I in its flexible form - 81.2%; type II in its rigid form - 0%; type II in its flexible form - 100%; type III in its rigid form - 47.6%; and type III in its flexible form - 100%. Of the 93 fingers included in this study, 42% abandoned the treatment. Conclusion Static orthoses are a safe alternative to surgical procedures, with low execution complexity for camptodactyly treatment.

Strategy for the Conservative Treatment of Type-III Camptodactyly in Children with Beals-Hecht Syndrome.

The authors present a successful case in the conservative treatment of type-III camptodactyly in a patient with Beals-Hecht syndrome. Camptodactyly is a flexion deformity of the proximal interphalangeal (PIP) joint, in the anteroposterior direction, painless and bilateral in 2/3 of the cases. Type-III is the most severe and disabling form, as it usually affects several fingers and is associated with syndromes and other malformations. The case herein reported had the correction achieved with the systematic use of static orthoses started at 7 months of age and completed after 23 and a half months of the intervention.

BMP-4, TGF-ß and Smad3 as Modulators of Synovial Fluid Cells Viability.

Objective Our goal was to evaluate the modulation of the synovial fluid cells (SFC) from patients with and without osteoarthritis (OA) by bone morphogenetic protein 4 (BMP-4), Smad-3 and transforming growth factor beta (TGF-ß). Methods Synovial fluid was collected from patients submitted to knee arthroscopy or replacement and were centrifuged to isolate cells from the fluid. Cells were cultured for 21 days and characterized as mesenchymal stem cells (MSCs) according to the criteria of the International Society of Cell Therapy. Then, we performed an [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay (MTT) assay after exposing cells with and without OA to TGF-ß, Smad3 and BMP-4 pathway inhibitors and to different concentrations of BMP4. Results Exposure to the TGF-ß, Smad3 and BMP-4 inhibitors modifies the mitochondrial activity of the SFCs. The activity of the SFCs is modified by influences of increasing concentrations of BMP4, but there is no difference in cellular activity between patients with and without OA. Conclusion TGF-ß, Smad3 and BMP-4 modulate the activity of SFCs from patients with and without knee OA.

Lateral Gradual Opening Osteotomy of the Tibia With Monolateral External Fixator for Correcting the Valgus Deformity of the Tibia.

The valgus deformity in the tibia requires correction because it places increased pressure on the lateral compartment of the knee, intensifying the degenerative process. Correction strategies are diverse and depend on patient profile, age, and soft-tissue conditions as well as the orthopaedic surgeon's experience with different surgical materials. Deformity size and location are the primary factors contributing to material and shape choice, whether gradual or acute. The only gradual correction approach involves the use of a monolateral or circular external fixator. This is the only indication for correction in cases of excellent deformity, soft-tissue involvement, and a history of bone infection. This study aimed to present a gradual correction technique for tibial valgus deformity using a monolateral external fixator as well as its postoperative follow-up. This technique has the advantages of greater patient acceptance, lighter assembly, and briefer distraction owing to the use of a single piece as well as the ability of the operated limb to bear a load the day after the surgical procedure and dynamic outpatient follow-up.

Harvest, Transport, and Storage of Fresh Humeral Head Osteochondral Allograft: Step-by-Step Protocol.

Articular cartilage defects are not common in the glenohumeral joint and are mostly found in patients after shoulder trauma, in patients with recurrent instability, or in patients who underwent previous surgical treatment. Articular cartilage defects lead to pain and loss of motion, consequently causing shoulder function impairment and reducing quality of life. In young patients, the use of osteochondral allografts for the treatment of humeral head defects may avoid well-known complications of shoulder arthroplasty. The goal of this Technical Note is to describe a step-by-step protocol for the harvesting, transport, and preservation of fresh humeral head osteochondral tissue for use in allograft transplantation.

Femoral torsion evaluation by computed tomography in a young Brazilian population with hip pain and femoroacetabular impingement.

INTRODUCTION: The aim of the study was to evaluate femoral torsion (FT) by computed tomography (CT) in young patients with hip pain and femoroacetabular impingement (FAI) in a Brazilian population. After the diagnosis of impingement, the complete analysis from the hip biomechanics and morphology has become essential. METHODS: Forty-one patients from 18 to 45 years presenting hip pain without arthrosis (Tönnis<2) were evaluated by CT scan from February 2017 to February 2018. All patients presented hip pain for at least 3 months and FAI. They have followed the same radiographic protocol and modified Harris Hip Score (mHHS) evaluation. Statistical analyses by software R version 3.4.4. with significance p < 0,05. RESULTS: After exclusion criterias, twenty-six patients (9 bilateral) were included, mostly man (73%). The average age was 35 years for both genders. BMI was 19 kg/m2 for women and 24 kg/m2 for men and the mean modified Harris Hip Score was 67 points. We have found femoral torsion changes in 11 hips with pain (31%) and high variability (60%). The mean FT was 14,5°, ranging from 0 to 39°. Patients with hip pain, CAM impingement and altered FT had no correlation when compared to controls (without pain) (p = 0.234), neither Mixed impingement (p = 0.314). Patients with Pincer impingement and painful hips had higher FT (16,63°) than controls (11,77°) (p = 0.045). Conclusion: The presence of torsional alterations in almost 1/3 of the patients with FAI and the high variability reveal the importance of measuring FT at this disease.

Protocol for Harvest, Transport and Storage of Human Osteochondral Tissue.

Objective To elaborate a protocol for the harvest, transport, and preservation of human osteochondral tissue for use in tissue banks (TBs). Methods Osteochondral fragments measuring 2 cm 3 of 5 corpse donors aged between 15 and 45 years old were analyzed. The samples were stored in cell preservation medium containing: human albumin, Iscove's and vancomycin preserved at 4°C. The concentration of proteoglycans in the extracellular medium was quantified by the use of Safranin-O, while tissue structural analysis was assessed by histological study with hematoxylin-eosin stained slides. The images obtained were analyzed according to the histological scores of Mankin and the score proposed by the OsteoArthritis Research Society International. The samples were analyzed with 0, 15, 30 and 45 days of preservation. Results The osteochondral fragments studied showed a progressive decrease in proteoglycan concentration with increased preservation time. After 30 days of preservation, structural changes were identified with discontinuity of the cartilage surface layer. According to the results obtained by the Mankin score, there was a statistically significant difference between 15 and 30 days of tissue preservation. Conclusion The protocol described defined knee transport immersed in Lactated Ringer at a controlled temperature of 10° C until its arrival at the TB. After processing, the preservation solution was composed of Iscove's serum-free cell culture medium supplemented with 10% human albumin and 100 µg/ml vancomycin. The tissue was preserved at a temperature of 4°C until the moment of transplantation characterizing the fresh preservation.

Viscosupplementation improves pain, function and muscle strength, but not proprioception, in patients with knee osteoarthritis: a prospective randomized trial.

OBJECTIVES: This study aimed to evaluate the clinical outcomes of intra-articular infiltration with hyaluronic acid and dexamethasone alone and in combination in the treatment of knee osteoarthritis (OA). METHOD: This prospective randomized trial evaluated 44 patients undergoing treatment for OA. Patients were selected through clinical and radiological analysis using the American College of Rheumatology criteria. We included patients aged between 50 and 70 years who presented with K-L stage ≤2 knee OA and normal limb alignment. Patients with a previous history of knee injury (ligamentous, meniscal or traumatic), infection, patellofemoral OA or chondroprotective drug use in the previous year were excluded. Participants were randomized into 3 groups and underwent treatment with viscosupplementation (VS, n=16), viscosupplementation plus dexamethasone (VD, n=16) or dexamethasone (DX, n=12). All patients were evaluated before and 6 weeks, 3 months and 6 months after infiltration. Analysis included a physical examination, the Western Ontario and McMaster Universities Arthritis Index (WOMAC) questionnaire (total score and domain subscores) and an evaluation of knee extensor and flexor strength and proprioception using an isokinetic dynamometer. RESULTS: VS significantly improved the WOMAC total score and subscores for pain, stiffness and function for up to 6 months after infiltration. It also improved knee extensor and flexor strength during the same period. Proprioception was not affected by any of the treatments. CONCLUSIONS: VS alone improved pain, stiffness and function according to the WOMAC total score and subscores and improved knee extensor and flexor strength, but not proprioception, for up to six months after infiltration. These findings suggest that VS has a positive effect on quadriceps arthrogenic inhibition.

Osteoarthritic Synovial Fluid Modulates Cell Phenotype and Metabolic Behavior In Vitro.

Synovial fluid holds a population of mesenchymal stem cells (MSC) that could be used for clinical treatment. Our goal was to characterize the inflammatory and metabolomic profile of the synovial fluid from osteoarthritic patients and to identify its modulatory effect on synovial fluid cells. Synovial fluid was collected from non-OA and OA patients, which was centrifuged to isolate cells. Cells were cultured for 21 days, characterized with specific markers for MSC, and exposed to a specific cocktail to induce chondrogenic, osteogenic, and adipogenic differentiation. Then, we performed a MTT assay exposing SF cells from non-OA and OA patients to a medium containing non-OA and OA synovial fluid. Synovial fluid from non-OA and OA patients was submitted to ELISA to evaluate BMP-2, BMP-4, IL-6, IL-10, TNF-α, and TGF-ß1 concentrations and to a metabolomic evaluation using 1H-NMR. Synovial fluid cells presented spindle-shaped morphology in vitro. Samples from OA patients formed a higher number of colonies than the ones from non-OA patients. After 21 days, the colony-forming cells from OA patients differentiated into the three mesenchymal cell lineages, under the appropriated induction protocols. Synovial fluid cells increased its metabolic activity after being exposed to the OA synovial fluid. ELISA assay showed that OA synovial fluid samples presented higher concentration of IL-10 and TGF-ß1 than the non-OA, while the NMR showed that OA synovial fluid presents higher concentrations of glucose and glycerol. In conclusion, SFC activity is modulated by OA synovial fluid, which presents higher concentration of IL-10, TGF-ß, glycerol, and glucose.

Strategy for the Conservative Treatment of Type-III Camptodactyly in Children with Beals-Hecht Syndrome / Estratégia para o tratamento conservador da camptodactilia de tipo III em criança com síndrome de Beals-Hecht

Abstract The authors present a successful case in the conservative treatment of type-III camptodactyly in a patient with Beals-Hecht syndrome. Camptodactyly is a flexion deformity of the proximal interphalangeal (PIP) joint, in the anteroposterior direction, painless and bilateral in 2/3 of the cases. Type-III is the most severe and disabling form, as it usually affects several fingers and is associated with syndromes and other malformations. The case herein reported had the correction achieved with the systematic use of static orthoses started at 7 months of age and completed after 23 and a half months of the intervention.

Resumo Os autores apresentam um caso bem-sucedido no tratamento conservador da camptodactilia de tipo III em paciente com síndrome de Beals-Hecht. A camptodactilia é uma deformidade em flexão da articulação interfalangeana proximal (IFP), no sentido anteroposterior, indolor, e bilateral em 2/3 dos casos. A de tipo III é a forma mais grave e incapacitante, pois geralmente acomete vários dedos e está associada a síndromes e outras malformações. O caso apresentado teve a correção alcançada com o uso sistemático de órteses estáticas iniciado aos 7 meses de idade e concluído após 23 meses e meio de intervenção.

Chronic Periodontitis and RANKL/OPG Ratio in Peri-Implant Mucosae Inflammation.

tHistory of chronic periodontitis (CP) is a risk factor for oseointegration failure. The osteoclastogenesis system (RANK, RANKL and OPG) is critical for bone homeostatic control. We investigated the levels of OPG and RANKL in peri-implant tissues from volunteers with and without a history of CP and their association with mucosae inflammation. This is a single-blind case-contro study. Diagnosis of a history of CP and peri-implant examination was performed on 46 volunteers, divided into control (without history of CP, n=26) and CP group (with history of CP, n=20). Gingival biopsies were harvested during implant exposure. Quantitative PCR evaluated OPG/RANKL mRNA expressions. OPG and RANKL proteins were analyzed by western blot and immunohistochemistry assay. The chi-square test analyzed the significance of nominal variables between groups while continuous variables were analyzed by T-test or Mann-Whitney test, after Shapiro-Wilk test evaluation. The 2-ΔΔCT Livak method calculation evaluated the gene expression. Values of p<0.05 were considered statistically significant. Volunteers with CP history had 23 times higher chance of developing mucosae inflammation. High mucosae levels of RANKL (p=0.04) and RANKL/OPG (p=0.001) mRNA expressions were observed in CP group. CP volunteers showed increased RANKL protein levels in opposition to decreased OPG expression. Even without active periodontitis, volunteers with a history of CP had elevated gingival levels of RANKL/OPG and higher correlation with peri-implant mucosae inflammation and implant loss.

BMP-4, TGF-β and Smad3 as Modulators of Synovial Fluid Cells Viability / BMP-4, TGF-β e Smad3 como moduladores da viabilidade das células do líquido sinovial

Abstract Objective Our goal was to evaluate the modulation of the synovial fluid cells (SFC) from patients with and without osteoarthritis (OA) by bone morphogenetic protein 4 (BMP-4), Smad-3 and transforming growth factor beta (TGF-β). Methods Synovial fluid was collected from patients submitted to knee arthroscopy or replacement and were centrifuged to isolate cells from the fluid. Cells were cultured for 21 days and characterized as mesenchymal stem cells (MSCs) according to the criteria of the International Society of Cell Therapy. Then, we performed an [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay (MTT) assay after exposing cells with and without OA to TGF-β, Smad3 and BMP-4 pathway inhibitors and to different concentrations of BMP4. Results Exposure to the TGF-β, Smad3 and BMP-4 inhibitors modifies the mitochondrial activity of the SFCs. The activity of the SFCs is modified by influences of increasing concentrations of BMP4, but there is no difference in cellular activity between patients with and without OA. Conclusion TGF-β, Smad3 and BMP-4 modulate the activity of SFCs from patients with and without knee OA.

Resumo Objetivo Nosso objetivo foi avaliar a modulação das células do líquido sinovial (SFCs, na sigla em inglês) de pacientes com e sem osteoartrite (OA) por proteína morfogenética óssea 4 (BMP-4), Smad3 e transformador do fator de crescimento β (TGF-β). Métodos O do líquido sinovial foi coletado de pacientes submetidos a artroscopia ou artroplastia do joelho, e centrifugados para isolar as células do liquido sinovial. As células foram cultivadas por 21 dias e caracterizadas como células-tronco mesenquimais (MSCs, na sigla em inglês) de acordo com os critérios da International Society of Cell Therapy. Em seguida, realizamos um ensaio de brometo de 3-4,5-dimetil-tiazol-2-il-2,5difeniltetrazólio (MTT) depois de expor células com e sem OA para TGF-β, inibidores de via Smad3 e BMP-4 e para diferentes concentrações de BMP-4. Resultados A exposição aos inibidores TGF-β, Smad3 e BMP-4 modifica a atividade mitocondrial das SFCs. A atividade das SFCs é modificada por influências sobre o aumento das concentrações de BMP-4, mas não há diferença na atividade celular entre pacientes com e sem OA. Conclusão TGF-β, Smad3 e BMP-4 modulam a atividade das SFCs de pacientes com e sem OA do joelho.

Vertical femoral neck fractures in young adults: a closed fixation strategy using a transverse cancellous lag screw.

Vertical femoral neck fractures (Pauwels type III classification) in young adults generally occur as a consequence of high-energy trauma and are frequently seen in association with multiple injuries. Considering the controversies regarding the optimal fixation for this fracture, our aim was to evaluate the clinical outcome of a closed fixation strategy for vertical femoral neck fractures in young adults using two parallel and one transverse cancellous lag screws. This was a single-surgeon, prospective study including 20 young adults with average age of 38.75 years (range 18-59 years) with a high-energy Pauwels III femoral neck fracture. Closed reduction and internal fixation with three cancellous lag screws were performed. The first screw was inserted crosswise to avoid further shear forces. Second and third parallel screws were placed above the lesser trochanter and centrally on the greater trochanter, respectively. Clinical outcomes were assessed by comparing postoperative and final follow-up radiographs 24 months post-injury. Eleven patients had an isolated vertical femoral neck fracture. Of these, five had further femoral neck comminution. Nine patients had an associated ipsilateral femoral shaft fracture. All fractures were displaced at the time of the first radiological evaluation. Closed reduction quality was considered excellent or good in 15 patients. After 24 months, bone union was achieved in 16 cases. Osteonecrosis of the femoral head developed in association with two fractures, and a nonunion developed in association with two fractures. We conclude that vertical high-energy femoral neck fractures can be treated successfully with internal fixation with two parallel cancellous lag screws positioned above the lesser trochanter and a third screw inserted centrally on the greater trochanter at an angle perpendicular to the fracture line.

Liposclerosing myxofibrous tumor: A series of 9 cases and review of the literature.

BACKGROUND: Liposclerosing myxofibrous primary bone tumor is a rare benign bone lesion that was characterized by complex mixture of various histological elements. METHODS: We have studied the radiological, clinical and pathological features of nine patients with this disorder. Pain and limping were the main symptoms. RESULTS: Radiographic images typically showed a geographic lytic lesion with thick sclerotic margin, reflecting a pattern of slow growth. Histological sections revealed a polymorphic neoplasia characterized by predominant proliferation of stellate and fusiform cells aimed the myxoid matrix. CONCLUSIONS: These features suggest that the lesion may represent a variant of fibrous dysplasia with a high risk of malignant transformation.

Mesenchymal stromal cells from bone marrow treated with bovine tendon extract acquire the phenotype of mature tenocytes.

OBJECTIVE: This study evaluated in vitro differentiation of mesenchymal stromal cells isolated from bone marrow, in tenocytes after treatment with bovine tendon extract. METHODS: Bovine tendons were used for preparation of the extract and were stored at -80 °C. Mesenchymal stromal cells from the bone marrow of three donors were used for cytotoxicity tests by means of MTT and cell differentiation by means of qPCR. RESULTS: The data showed that mesenchymal stromal cells from bone marrow treated for up to 21 days in the presence of bovine tendon extract diluted at diminishing concentrations (1:10, 1:50 and 1:250) promoted activation of biglycan, collagen type I and fibromodulin expression. CONCLUSION: Our results show that bovine tendon extract is capable of promoting differentiation of bone marrow stromal cells in tenocytes.

OBJETIVO: O estudo avalia a diferenciação in vitro das células mesenquimais isoladas do estroma da medula óssea em tenócitos após tratamento com extrato de tendão bovino. MÉTODOS: Tendões bovinos foram usados para confecção do extrato e estocados a −80 °C. Células mesenquimais do estroma da medula óssea (BMSCs) de três doadores foram usadas para os testes de citotoxicidade por MTT e diferenciação celular por qPCR. RESULTADOS: Os dados mostram que células mesenquimais do estroma da medula óssea tratadas por até 21 dias em presença do extrato de tendão bovino diluído em concentrações crescentes (1:10, 1:50 e 1:250) promovem a ativação da expressão de biglican, colágeno tipo I e fibromodulina. CONCLUSÃO: Nossos resultados mostram que o extrato de tendão bovino é capaz de promover a diferenciação das BMSCs em tenócitos.

Vascular Endothelial Growth Factor Receptor-2 Polymorphisms Have Protective Effect against the Development of Tendinopathy in Volleyball Athletes.

The aim of the study was to investigate whether genetic variants in VEGF and KDR genes can be correlated with susceptibility of tendinopathy in volleyball athletes. This study was conducted at the Brazilian Volleyball Federation, and comprised 179 volleyball athletes: 88 had a confirmed diagnosis of tendinopathy (cases), whereas 91 had no evidence of the disease (controls). The VEGF (-2578C>A, -460T>C and +936C>T) and KDR (-604C>T, 1192G>A and 1719T>A) polymorphisms were determined by TaqMan real-time polymerase chain reaction. The odds ratio (OR) with their 95% confidence intervals (CI) were calculated using an unconditional logistic regression model. The evaluation of demographic and clinical characteristics revealed the athlete age (P < 0.001), years of practice in volleyball (P < 0.001) and presence of pain (P = 0.001) were risk factors for tendinopathy. KDR 1192 GA and GA + AA genotypes were associated with lower risk of tendinopathy (OR: 0.41, 95% CI: 0.19-0.88 and OR: 0.47, 95% CI: 0.23-0.98, respectively). The KDR (-604C>T, 1192G>A and 1719T>A) haplotypes CGA and CAT were associated with decreased tendinopathy risk (OR: 0.46, 95% CI: 0.21-0.99 and OR: 0.23, 95% CI: 0.07-0.76, respectively). With regards to pain, traumatic lesion and away from training due to injury, VEGF and KDR polymorphisms were not associated with clinical symptoms complaints. The present results provide evidence that the KDR polymorphisms were associated with development of tendinopathy, and can contribute to identify new therapeutic targets or personalized training programs to avoid tendinopathy development in athletes.

Protocol for Harvest, Transport and Storage of Human Osteochondral Tissue / Protocolo para captação, transporte e preservação de tecido osteocondral humano

Abstract Objective To elaborate a protocol for the harvest, transport, and preservation of human osteochondral tissue for use in tissue banks (TBs). Methods Osteochondral fragments measuring 2 cm3 of 5 corpse donors aged between 15 and 45 years old were analyzed. The samples were stored in cell preservation medium containing: human albumin, Iscove's and vancomycin preserved at 4ºC. The concentration of proteoglycans in the extracellular medium was quantified by the use of Safranin-O, while tissue structural analysis was assessed by histological study with hematoxylin-eosin stained slides. The images obtained were analyzed according to the histological scores of Mankin and the score proposed by the OsteoArthritis Research Society International. The samples were analyzed with 0, 15, 30 and 45 days of preservation. Results The osteochondral fragments studied showed a progressive decrease in proteoglycan concentration with increased preservation time. After 30 days of preservation, structural changes were identified with discontinuity of the cartilage surface layer. According to the results obtained by the Mankin score, there was a statistically significant difference between 15 and 30 days of tissue preservation. Conclusion The protocol described defined knee transport immersed in Lactated Ringer at a controlled temperature of 10º C until its arrival at the TB. After processing, the preservation solution was composed of Iscove's serum-free cell culture medium supplemented with 10% human albumin and 100 µg/ml vancomycin. The tissue was preserved at a temperature of 4ºC until the moment of transplantation characterizing the fresh preservation.

Resumo Objetivo Elaborar um protocolo para a captação, transporte e preservação de tecido osteocondral humano para utilização em banco de tecidos (BT). Métodos Foram analisados fragmentos osteocondrais com dimensão de 2 cm3 de 5 doadores cadáveres com idades entre 15 e 45 anos. As amostras foram armazenadas em meio de preservação celular contendo: albumina humana, Iscove's e vancomicina preservados à temperatura de 4ºC. A concentração de proteoglicanos no meio extracelular foi quantificada pelo uso de Safranina-O, enquanto a análise estrutural do tecido foi avaliada através de estudo histológico com lâminas coradas em hematoxilina-eosina. As imagens obtidas foram analisadas segundo os escore histológicos de Mankin e o escore proposto pela OsteoArthritis Research Society International. As amostras foram analisadas com 0, 15, 30 e 45 dias de preservação. Resultados Os fragmentos osteocondrais estudados apresentaram diminuição progressiva na concentração de proteoglicanos com o aumento do tempo de preservação. Após 30 dias de preservação, foram identificadas alterações estruturais com descontinuidade da camada superficial da cartilagem. Segundo os resultados obtidos pelo escore de Mankin, houve diferença com significância estatística entre 15 e 30 dias de preservação do tecido. Conclusão O protocolo descrito definiu o transporte de joelho em bloco imerso em Ringer Lactato em temperatura controlada a 10ºC até sua chegada ao BT. Após o processamento, a solução de preservação foi composta por meio de cultura celular sem soro Iscove's suplementado com albumina humana a 10% e vancomicina 100 µg/mL. O tecido foi preservado à temperatura de 4ºC até o momento do transplante caracterizando a preservação a fresco.