Efficiency analysis of commercial polymeric membranes for bone regeneration in rat cranial defects

Purpose: To evaluate the in vivo efficiency of commercial polymeric membranes for guided bone regeneration. Methods: Rat calvarial critical size defects was treated with LuminaCoat (LC), Surgitime PTFE (SP), GenDerm (GD), Pratix (PR), Techgraft (TG) or control (C-) and histomorphometric analysis determined the percentage of new bone, connective tissue and biomaterial at 1 or 3 months. Statistical analysis used ANOVA with Tukey's post-test for means at same experimental time and the paired Student's t test between the two periods, considering p < 0.05. Results: New bone at 1 month was higher for SP, TG and C-, at 3 months there were no differences, and between 1 and 3 months PR had greater increase growthing. Connective tissue at 1 month was higher for C-, at 3 months for PR, TG and C-, and between 1 and 3 months C- had sharp decline. Biomaterial at 1 month was higher for LC, in 3 months for SP and TG, and between 1 and 3 months, LC, GD and TG had more decreasing mean. Conclusion: SP had greater osteopromotive capacity and limitation of connective ingrowth, but did not exhibit degradation. PR and TG had favorable osteopromotion, LC less connective tissue and GD more accelerated biodegradation.

Influence of a novel scaffold composed of polyurethane, hydroxyapatite, and decellularized bone particles on the healing of fourth metacarpal defects in mares.

OBJECTIVE: To determine the effect of a novel scaffold, designed for use in bone regeneration, on healing of splint bone segmental defects in mares. STUDY DESIGN: In vivo experimental study. SAMPLE POPULATION: Five adult mares (4-10 years old; mean weight, 437.7 kg ± 29 kg). METHODS: Bilateral 2-cm full-thickness defects were created in the fourth metacarpal bones (MCIV) of each horse. Each defect was randomly assigned to either a novel scaffold treatment (n = 5) or an untreated control (n = 5). The scaffold was composed of polyurethane, hydroxyapatite, and decellularized bone particles. Bone healing was assessed for a period of 60 days by thermography, ultrasonography, radiography, and computed tomography (CT). Biopsies of each defect were performed 60 days after surgery for histological evaluation. RESULTS: On the basis of radiographic analysis, scaffold-treated defects had greater filling (67.42% ± 26.7%) compared with untreated defects (35.88% ± 32.7%; P = .006). After 60 days, CT revealed that the density of the defects treated with the scaffolds (807.80 ± 129.6 Hounsfield units [HU]) was greater than density of the untreated defects (464.80 ± 81.3 HU; P = .004). Evaluation of histology slides provided evidence of bone formation within an average of 9.43% ± 3.7% of the cross-sectional area of scaffolds in contrast to unfilled defects in which connective tissue was predominant throughout the biopsy specimens. CONCLUSION: The novel scaffold was biocompatible and supported bone formation within the MCIV segmental defects. CLINICAL SIGNIFICANCE: This novel scaffold offers an effective option for filling bone voids in horses when support of bone healing is indicated.

Cicatrização da musculatura reto-abdominal em coelhos submetidos à laparorrafia com fios de sutura à base de quitosana, categute cromado e poliglactina 910 / [Healing of the rectus abdominis muscle in rabbits submitted to laparorrhaphy with suture-based chitosan, chromic catgut and polyglactin 910]

Objetivou-se, com este estudo, avaliar o processo de cicatrização da musculatura reto-abdominal em coelhos submetidos à laparorrafia, utilizando-se o fio de sutura à base de quitosana, comparando-o aos fios de categute cromado e poliglactina 910. Foram utilizados 24 coelhos adultos, divididos aleatoriamente em quatro grupos: quitosana e categute 15 dias (QC-15dias), quitosana e categute 30 dias (QC-30 dias), quitosana e poliglactina 910 15 dias (QP-15 dias) e quitosana e poliglactina 910 30 dias (QP-30 dias). Cada grupo foi composto por seis coelhos, nos quais foram realizadas duas incisões, uma do lado direito e outra do lado esquerdo e, posteriormente, a laparorrafia, com o fio de quitosana de um lado e o categute cromado ou poliglactina 910 do outro. Realizou-se análise clínico-cirúrgica, histológica e avaliação de achados de necropsia, além de testes de citotoxicidade e de mecânica no fio de quitosana. Ele apresentou baixa resistência mecânica e citotóxica. O fio de quitosana não proporcionou uma cicatrização satisfatória em coelhos, pois desencadeou uma resposta inflamatória acentuada.(AU)

The objective of this study was to evaluate the healing process of the recto-abdominal muscles in rabbits submitted to laparorrhaphy using chitosan-based suture yarn, comparing it to chrome catgut and polyglactin 910 yarns. Twenty-four adult rabbits were divided in to four random groups: chitosan and polyglactin 910 15 days (QP-15 days) and chitosan and polyglactin 910 30 days (QC-30 days), chitosan and polyglactin 910 15 days (QP-15 days) QP-30 days). Each group consisted of six rabbits, in which two incisions were made, one on the right side and one on the left side, and later the laparorraphy with the chitosan yarn on one side and chromed catgut or polyglactin 910 on the other. Clinical-surgical, histological and necropsy findings were evaluated, as well as cytotoxicity and mechanical tests on the chitosan wire. It presented low mechanical and cytotoxic resistance. Chitosan thread did not provide satisfactory healing in rabbits, as it triggered a marked inflammatory response.(AU)

Membrana de látex natural de Hevea brasiliensis auxilia no processo de reparação tecidual em bovinos / Natural latex rubber from Hevea brasiliensis aids tissue repair of bovine

Feridas cutâneas em bovinos são um constante desafio clínico cirúrgico por desencadearem perdas econômicas bastante significativas. O látex proveniente da seiva da seringueira (Hevea brasiliensis) apresenta potencial terapêutico para incrementar o processo de reparação tecidual. Portanto, pretendeu-se com esse estudo avaliar o tipo de reação tecidual e os possíveis mecanismos de angiogênese desencadeados pelo implante de uma membrana de látex natural em bovinos. Para tal, foram utilizados seis bovinos da raça Nelore, submetidos ao implante subcutâneo experimental de três fragmentos de membranas de látex natural. Foram coletadas amostras de tecido e da membrana aos 15, 30 e 45 dias após a implantação, para avaliações histológicas, ultraestruturais por microscopia eletrônica de varredura e imunoistoquímicas com anticorpos antimarcador de macrófagos (MAC), CYR 61 e VEGF. O implante de látex proporcionou aumento da angiogênese e reparação tecidual em bovinos, não mediada pela expressão do VEGF e CYR 61.(AU)

Cattle wounds are a constant surgical and clinical challenge, leading to important economical losses. The latex from the sap of the rubber tree (Hevea brasiliensis) has therapeutic potential to enhance tissue repair process. Therefore, we evaluated the type of tissue reaction and possible mechanisms of angiogenesis triggered by implanting natural latex rubber in bovine species. Six Nelore bovines were subjected to subcutaneous experimental implant of three fragments of natural rubber latex membranes. Tissue and rubber membrane samples were harvested at 15, 30 and 45 days implantation for histology, scanning electron microscopy and immunohistochemical evaluation with anti macrophage marker (MAC), anti CYR 61, anti VEGF antibodies. The latex membrane estimulates tissue reaction and repair and significant angiogenesis stimuli without activating CYR 61 and VEGF pathways.(AU)

Treatment of periodontal disease with guided tissue regeneration technique using a hydroxyapatite and polycaprolactone membrane / Tratamento da doença periodontal pela técnica da regeneração tecidual guiada utilizando uma membrana de hidroxiapatita e policaprolactona

The aim of this study was to evaluate the use of a malleable membrane composed of hydroxyapatite (60%) and polycaprolactone (40%) as treatment of periodontal disease experimentally induced in dogs. A bone defect of standardized dimensions was created between the roots of the third and fourth premolar of 12 dogs for periodontal disease induction. Six dogs had the defect covered by the membrane and six dogs received only standard treatment for periodontal disease, also applied to dogs in the treated group. The animals were clinically monitored during the experiment. Radiographs were taken after surgery and at 60 days after treatment initiation. Clinical attachment level was also assessed in those moments. On the 60th day, dental sample of all animals, containing tooth, defect and periodontal tissues, were harvested, fixed in formalin and analyzed by microtomography and histology. During the experimental period, the animals showed no pain and purulent discharge, however, there was dehiscence in 50% of animals and membrane exposure in five out of six animals in the treated group. Clinical attachment level showed no difference between groups. Radiographs showed radiopacity equal to the alveolar bone in both groups. The microtomography revealed that the control group had higher bone volume in the defect compared to the treated group; however, the furcation was not filled by new alveolar bone in any animal. Histological analysis revealed that junctional epithelium invasion was lighter in the control group. New bone was only observed in the apical edge of the defect in both groups. Although the composite is biocompatible and able to keep the space of the defect, it did not promote periodontal tissue regeneration within 60 days of observation.(AU)

O objetivo do presente trabalho foi avaliar a utilização de membrana moldável constituída por hidroxiapatita (60%) e policaprolactona (40%) como tratamento da doença periodontal, induzida experimentalmente em cães. Um defeito ósseo de dimensões padronizadas foi realizado entre as raízes do terceiro e do quarto pré-molares de 12 cães para indução da doença periodontal. Todos os cães receberam tratamento padrão para doença periodontal, e seis desses animais foram tratados também com a aplicação da membrana sobre o defeito. Os animais foram acompanhados clinicamente durante o experimento. Radiografias foram realizadas no pós-operatório e aos 60 dias após o início do tratamento. O nível clínico de inserção também foi avaliado nesses momentos. Aos 60 dias, a amostra dental de todos os animais contendo o dente, o defeito e os tecidos periodontais foi coletada, fixada em formol e analisada por microtomografia e histologia. Durante o período experimental, os animais não apresentaram dor e secreção purulenta, entretanto houve deiscência em 50% dos animais e exposição de membrana em cinco dos seis animais do grupo tratado. Nível clínico de inserção não apresentou diferença entre os grupos. As imagens radiográficas mostraram radiopacidade igual ao osso alveolar em ambos os grupos. A microtomografia revelou que o grupo controle apresentou maior volume ósseo no defeito em relação ao grupo tratado, no entanto, em todos os animais, a região de furca não foi preenchida por novo osso alveolar. A análise histológica revelou que a invasão por epitélio juncional foi mais discreta no grupo controle. Osso novo foi apenas observado na borda apical do defeito em ambos os grupos. Embora o compósito seja biocompatível e tenha sido capaz de manter o espaço do defeito, ele não promoveu a regeneração dos tecidos periodontais no período de 60 dias de observação.(AU)

Isolation, expansion and differentiation of mesenchymal stromal cells from rabbits' bone marrow / Isolamento, expansão e diferenciação de celulas tronco mesenquimais oriundas da medula óssea de coelhos

Tissue engineering has been a fundamental technique in the regenerative medicine field, once it permits to build tri-dimensional tissue constructs associating undifferentiated mesenchymal cells (or mesenchymal stromal cells - MSCs) and scaffolds in vitro. Therefore, many studies have been carried out using these cells from different animal species, and rabbits are often used as animal model for in vivo tissue repair studies. However, most of the information available about MSCs harvesting and characterization is about human and murine cells, which brings some doubts to researchers who desire to work with a rabbit model in tissue repair studies based on MSCs. In this context, this study aimed to add and improve the information available in the scientific literature providing a complete technique for isolation, expansion and differentiation of MSCs from rabbits. Bone marrow mononuclear cells (BMMCs) from humerus and femur of rabbits were obtained and to evaluate their proliferation rate, three different culture media were tested, here referred as DMEM-P, DMEM´S and α-MEM. The BMMCs were also cultured in osteogenic, chondrogenic and adipogenic induction media to prove their multipotentiality. It was concluded that the techniques suggested in this study can provide a guideline to harvest and isolate MSCs from bone marrow of rabbits in enough amount to allow their expansion and, based on the laboratory experience where the study was developed, it is also suggested a culture media formulation to provide a better cell proliferation rate with multipotentiality preservation.

A engenharia de tecidos tem sido uma técnica fundamental no campo da medicina regenerativa, uma vez que permite a criação de peças teciduais tri-dimensionais por meio da associação de células mesenquimais indiferenciadas (ou células estromais mesenquimais - CEMs) e moldes de biomateriais in vitro. Assim, muitos estudos têm sido realizados utilizando estas células oriundas de diferentes espécies animais, e os coelhos são frequentemente utilizados como um modelo animal para estudos in vivo de reparação tecidual. No entanto, a maioria das informações disponíveis sobre a coleta e caracterização de CEMs referem-se às células humanas e murinas, o que traz algumas dúvidas para pesquisadores que desejam trabalhar com coelhos em estudos de reparação de tecidos baseados em CEMs. Neste contexto, o presente estudo objetivou contribuir e aprimorar as informações disponíveis na literatura científica fornecendo uma técnica completa para o isolamento, expansão e diferenciação das MSCs de coelhos. Células mononucleares da medula óssea (CMMOs) do úmero e fêmur de coelhos foram obtidas e, para avaliar sua taxa de proliferação, três meios de cultura diferentes foram testadas, aqui referidos como DMEM-P, DMEM'S e α-MEM. As CMMOs também foram cultivadas em meios de indução osteogênico, condrogênico, e linhagens adipogênico para provar a sua multipotencialidade. Concluiu-se que as técnicas sugeridas neste estudo podem fornecer um guia para a coleta e isolamento de CEMs da medula óssea de coelhos em quantidade suficiente para permitir a sua expansão e, com base na experiência de laboratório onde o estudo foi desenvolvido, é também sugerida uma formulação de meio de cultivo para proporcionar uma melhor taxa de proliferação celular com preservação da multipotencialidade.

Mecanismos moleculares de reparación mediante la técnica Electrólisis Percutánea Intratisular en la tendinosis rotuliana / Molecular repair mechanisms using the Intratissue Percutaneous Electrolysis technique in patellar tendonitis

Objetivo. Investigar los mecanismos moleculares de respuesta tisular tras el tratamiento con la técnica Electrólisis Percutánea Intratisular (EPI®) en la tendinosis inducida por colagenasa tipo i en ratas Sprague Dawley. Métodos. En una muestra de 24 ratas Sprague Dawley de 7 meses de edad y 300 g se indujo tendinosis mediante la inyección en el tendón rotuliano de 50 μg de colagenasa tipo i . Se procedió a dividir la muestra en 4 grupos: un grupo control, un grupo colagenasa y 2 grupos de tratamiento con técnica EPI® a 3 y 6 mA, respectivamente. Se aplicó una sesión de tratamiento EPI® y tras 3 días se procedió al análisis de los tendones mediante técnicas de inmunodetección y electroforesis. Se analizaron las proteínas citocromo C, Smac/Diablo, factor de crecimiento endotelial vascular y su receptor 2. También se analizó el factor de transcripción nuclear peroxisoma proliferador activado del receptor gamma. Resultados. Se observó un aumento estadísticamente significativo en la expresión del citocromo C, Smac/Diablo, factor de crecimiento endotelial vascular, su receptor 2 y peroxisoma proliferador activado del receptor gamma en los grupos a los que se les aplicó la técnica EPI® respecto al grupo control. Conclusiones. La técnica EPI® produce, en la lesión tendinosa inducida con colagenasa tipo i en ratas, un aumento de los mecanismos moleculares antiinflamatorios y angiogénicos (AU)

Objective. To investigate the molecular mechanisms of tissue response after treatment with the Intratissue Percutaneous Electrolysis (EPI®) technique in collagenase-induced tendinopathy in Sprague-Dawley rats. Methods. Tendinopathy was induced by injecting 50 μg of type i collagenase into the patellar tendon of 24 Sprague Dawley rats of 7 months of age and weighting 300 g. The sample was divided into 4 groups: the control group, collagenase group, and two EPI® technique treatment groups of 3 and 6 mA, respectively. An EPI® treatment session was applied, and after 3 days, the tendons were analysed using immunoblotting and electrophoresis techniques. An analysis was also made of cytochrome C protein, Smac/Diablo, vascular endothelial growth factor and its receptor 2, as well as the nuclear transcription factor peroxisome proliferator-activated receptor gamma. Results. A statistically significant increase, compared to the control group, was observed in the expression of cytochrome C, Smac/Diablo, vascular endothelial growth factor, its receptor 2 and peroxisome proliferator-activated receptor gamma in the groups in which the EPI® technique was applied. Conclusions. EPI® technique produces an increase in anti-inflammatory and angiogenic molecular mechanisms in collagenase-induced tendon injury in rats (AU)

Tendon regeneration in human and equine athletes: Ubi Sumus-Quo Vadimus (where are we and where are we going to)?

Tendon injuries are one of the most common orthopaedic problems in both human and equine athletes. When a damaged tendon heals naturally, it loses a substantial part of the original strength and elasticity. Therefore, tendons recover structurally (reparation) but not functionally (regeneration) after conservative medical or surgical treatment. Since the structure and matrix composition of human and equine tendons share many similarities, the nature of tendon injuries are also strongly comparable in both species. Therefore, the evaluation of regenerative therapies in horses may have applications for future human medicine and vice versa. The current review focuses briefly on the physiology of human and equine tendon in order to better comprehend the modus operandi of this structure under pathophysiological circumstances. In addition, the reparative effects of conservative medical and surgical interventions are discussed concisely, and an extensive overview is given on the regenerative therapies that are currently being explored. For the latter, the results of equine clinical studies might prove invaluable for gaining additional insights into the treatment of human tendinopathies, since not all of these novel regenerative therapies have been evaluated in humans yet.

[Regenerative therapy for tendon and ligament disorders in horses. Terminology, production, biologic potential and in vitro effects]. / Regenerative Therapie von Sehnen- und Banderkrankungen bei Pferden. Terminologie, Herstellung, biologisches Potenzial und In-vitro-Effekte.

Conventional treatments of equine tendon injuries lead to an unsatisfactory healing process that usually results in a relatively high recurrence rate. Therefore, in recent years so-called regenerative therapeutics were studied scientifically in vitro and in laboratory animals. These include substances that ideally lead to the formation of replacement tissue, which in contrast to the low quality scar, has similar functional properties as the original intact tendon. Currently, a plethora of different substrates is either commercially available or can be produced in practice with the help of kits. The current knowledge on the production and the regenerative potential of nucleated cells like stem cells from bone marrow and fat tissue, of the blood products PRP (platelet rich plasma), ACP (autologous conditioned plasma), ACS (autologous conditioned serum) and of the scaffold substance UBM (urinary bladder matrix) are presented. Finally, the potential of some growth factors and of gene therapy is considered. Currently, it is assumed that the regeneration of tendon tissue is promoted by a complex interaction of scaffolds, growth factors and cells. At present, only very few studies are available which allow a comparison between these substances. Studies on the effect of regenerative substrates on tendons in live horses are presented elsewhere.

Tissue engineering in wound repair: the three "R"s--repair, replace, regenerate.

Horses are predisposed to traumatic wounds that can be labor intensive and expensive to manage. Skin has a considerable potential for efficient and functional repair however, while cutaneous repair is a regenerative process in the fetus, this capability declines in late gestation as inflammation and scarring alter the outcome of healing. The historical gold standard for replacement of lost skin is the autologous skin graft. However, the horse's lack of redundant donor skin limits the practicality of full-thickness grafting to smaller wounds; moreover, graft failure is relatively common in equine patients as a result of infection, inflammation, fluid accumulation beneath the graft, and motion. Tissue engineering has emerged as an interdisciplinary field with the aim to regenerate new biological material for replacing diseased or damaged tissues or organs. In the case of skin, the ultimate goal is to rapidly create a construct that effects the complete regeneration of functional skin, including all its layers and appendages. Moreover, an operational vascular and nervous network, with scar-free integration within the surrounding host tissue, is desirable. For this to be achieved, not only is an appropriate source of cells required, but also a scaffold designed from natural or synthetic polymers. The newly created tissue might finally meet the numerous needs and expectations of practitioners and surgeons managing a catastrophic wound in a horse.

Guided bone regeneration with beta-tricalcium phosphate and poly L-lactide-co-glycolide-co-epsilon-caprolactone membrane in partial defects of canine humerus.

This study was performed to evaluate the effect of betatricalcium phosphate and poly L-lactide-co-glycolide-coepsilon- caprolactone (TCP/PLGC) membrane in the repair of partial bone defects in canine proximal humerus. Three adult mixed-breed dogs were used during the experimental period. The length of the defect was quarter of the full length of humerus, and width of the defect was quarter of middle diameter of the lateral aspect of humerus. The humeri of each dog were divided into treatment (TCP/ PLGC) and control groups. The defect was covered with TCP/PLGC membrane in treatment group. To evaluate regeneration of the bone, computerized tomography (CT) and histopathologic examination were performed. The radiopaque lines were appeared at the original defect sites in TCP/PLGC group but below the original site in control at 4th week. Radiopacity and thickness of the defect sites, and radiopaque lines were more increased at 8th week than those of 4th week. Histopathologic findings revealed fibrous connective tissue migration into the defect and the migration inhibited the structure of new cortex to be placed in the original level in control whereas new cortex growth was found in the level of original line in TCP/ PLGC group. However, the new cortical bone in the TCP/ PLGC group was thinner and less organized than the adjacent intact cortex, and the amount of new cancellous bones were also scanty. The result suggested that TCP/ PLGC membrane is a good guided bone regeneration material to restore the original morphology of humerus in partial defect.

Alteración periodontal en un perro de 6 años: tratamiento mediante regeneración tisular guiada para disminuir la movilidad y estabilizar piezas / No disponible

No disponible