Research progress in leveraging biomaterials for enhancing NK cell immunotherapy / 浙江大学学报·医学版

NK cell immunotherapy is a promising antitumor therapeutic modality after the development of T cell immunotherapy. Structural modification of NK cells with biomaterials may provide a precise, efficient, and low-cost strategy to enhance NK cell immunotherapy. The biomaterial modification of NK cells can be divided into two strategies: surface engineering with biomaterials and intracellular modification. The surface engineering strategies include hydrophobic interaction of lipids, receptor-ligand interaction between membrane proteins, covalent binding to amino acid residues, click reaction and electrostatic interaction. The intracellular modification strategies are based on manipulation by nanotechnology using membranous materials from various sources of NK cells (such as exosome, vesicle and cytomembranes). Finally, the biomaterials-based strategies regulate the recruitment, recognition and cytotoxicity of NK cells in the solid tumor site in situ to boost the activity of NK cells in the tumor. This article reviews the recent research progress in enhancing NK cell therapy based on biomaterial modification, to provide a reference for further researches on engineering NK cell therapy with biomaterials.

Spatiotemporal cellular dynamics and molecular regulation of tooth root ontogeny / 国际口腔科学杂志·英文版

Tooth root development involves intricate spatiotemporal cellular dynamics and molecular regulation. The initiation of Hertwig's epithelial root sheath (HERS) induces odontoblast differentiation and the subsequent radicular dentin deposition. Precisely controlled signaling pathways modulate the behaviors of HERS and the fates of dental mesenchymal stem cells (DMSCs). Disruptions in these pathways lead to defects in root development, such as shortened roots and furcation abnormalities. Advances in dental stem cells, biomaterials, and bioprinting show immense promise for bioengineered tooth root regeneration. However, replicating the developmental intricacies of odontogenesis has not been resolved in clinical treatment and remains a major challenge in this field. Ongoing research focusing on the mechanisms of root development, advanced biomaterials, and manufacturing techniques will enable next-generation biological root regeneration that restores the physiological structure and function of the tooth root. This review summarizes recent discoveries in the underlying mechanisms governing root ontogeny and discusses some recent key findings in developing of new biologically based dental therapies.

Role of dendritic cells in MYD88-mediated immune recognition and osteoinduction initiated by the implantation of biomaterials / 国际口腔科学杂志·英文版

Bone substitute material implantation has become an important treatment strategy for the repair of oral and maxillofacial bone defects. Recent studies have shown that appropriate inflammatory and immune cells are essential factors in the process of osteoinduction of bone substitute materials. Previous studies have mainly focused on innate immune cells such as macrophages. In our previous work, we found that T lymphocytes, as adaptive immune cells, are also essential in the osteoinduction procedure. As the most important antigen-presenting cell, whether dendritic cells (DCs) can recognize non-antigen biomaterials and participate in osteoinduction was still unclear. In this study, we found that surgical trauma associated with materials implantation induces necrocytosis, and this causes the release of high mobility group protein-1 (HMGB1), which is adsorbed on the surface of bone substitute materials. Subsequently, HMGB1-adsorbed materials were recognized by the TLR4-MYD88-NFκB signal axis of dendritic cells, and the inflammatory response was activated. Finally, activated DCs release regeneration-related chemokines, recruit mesenchymal stem cells, and initiate the osteoinduction process. This study sheds light on the immune-regeneration process after bone substitute materials implantation, points out a potential direction for the development of bone substitute materials, and provides guidance for the development of clinical surgical methods.

Research advances on biomaterials for the delivery of growth factors to regulate wound repair / 中华烧伤杂志

Wound repair is a highly coordinated and mutually regulated complex process involving various kinds of cells, extracellular matrices and cytokines. A variety of growth factors play an important regulatory role in wound healing, and it is critical to achieve effective delivery and sustained function of growth factors. In recent years, the application of biomaterials in tissue engineering has shown great potential, and the effective delivery of growth factors by biomaterials has attracted increasing attention. Based on this, this paper introduces the mechanism of related growth factors in the process of wound healing, focusing on the recent progress of biomaterial delivery of growth factors to accelerate wound healing, in order to provide new enlightenment for clinical wound treatment.

Biodegradation properties of multi-laminated small intestinal submucosa / 北京大学学报(医学版)

OBJECTIVE@#To study the biodegradation properties of multi-laminated small intestinal submucosa (mSIS) through in vitro and in vivo experiments, comparing with Bio-Gide, the most widely used collagen membrane in guided bone regeneration (GBR) technique, for the purpose of providing basis to investigate whether mSIS meets the requirements of GBR in dental clinics.@*METHODS@#The degradation properties were evaluated in vitro and in vivo. In vitro degradation was performed using prepared collagenase solution. Morphology of mSIS and Bio-Gide in degradation solution were observed and the degradation rate was calculated at different time points. In in vivo experiments, nine New Zealand rabbits were used for subcutaneous implantation and were divided into three groups according to observation intervals. Six unconnected subcutaneous pouches were made on the back of each animal and were embedded with mSIS and Bio-Gide respectively. At the end of weeks 4, 8, and 12 after operation, gross observation and HE staining were used to evaluate the degree of degradation and histocompatibility.@*RESULTS@#In vitro degradation experiments showed that mSIS membrane was completely degraded at the end of 12 days, while Bio-Gide was degraded at the end of 7 days. Besides, mSIS maintained its shape for longer time in the degradation solution than Bio-Gide, indicating that mSIS possessed longer degradation time, and had better ability to maintain space than Bio-Gide. In vivo biodegradation indicated that after 4 weeks of implantation, mSIS remained intact. Microscopic observation showed that collagen fibers were continuous with a few inflammatory cells that infiltrated around the membrane. Bio-Gide was basically intact and partially adhered with the surrounding tissues. HE staining showed that collagen fibers were partly fused with surrounding tissues with a small amount of inflammatory cells that infiltrated as well. Eight weeks after operation, mSIS was still intact, and was partly integrated with connective tissues, whereas Bio-Gide membrane was mostly broken and only a few residual fibers could be found under microscope. Only a small amount of mSIS debris could be observed 12 weeks after surgery, and Bio-Gide could hardly be found by naked eye and microscopic observation at the same time.@*CONCLUSION@#In vitro degradation time of mSIS is longer than that of Bio-Gide, and the space-maintenance ability of mSIS is better. The in vivo biodegradation time of subcutaneous implantation of mSIS is about 12 weeks and Bio-Gide is about 8 weeks, both of which possess good biocompatibility.

Imunoproteção de ilhotas pancreáticas microencapsuladas em biomateriais inovadores e seu potencial terapêutico no diabetes mellitus tipo 1 / Immunoprotection of pancreatic islets microencapsulated in inovative biomaterials and its therapeutic potential in type 1 Diabetes Mellitus

O transplante de ilhotas microencapsuladas constitui uma alternativa terapêutica interessante para o Diabetes Mellitus tipo 1, permitindo um melhor controle glicêmico e eliminando a necessidade de imunossupressão. Entretanto, a manutenção a longo prazo da viabilidade das células-ß ainda é um desafio. No isolamento, a perda da matriz extracelular e as condições hipóxicas subsequentes afetam decisivamente a sobrevivência e funcionalidade das ilhotas. Objetivo Para diminuir o estresse sobre o enxerto, levando a um sucesso prolongado do transplante, propôs-se a adição de perfluorocarbono (PFC) ou laminina (LN), moléculas associadas respectivamente à oxigenação e interações célula-célula, ao biomaterial baseado em alginato, Biodritina, adequado ao encapsulamento celular. Metodologia Para testar a estabilidade das formulações PFC-Biodritina e LN-Biodritina, microcápsulas foram submetidas a diferentes estresses (rotacional, osmótico, temperatura e cultura) por 7 e 30 dias. A pureza do biomaterial foi avaliada pela coincubação com macrófagos murinos RAW264.7, por 3, 9 e 24h, quando a ativação dos macrófagos foi observada pela expressão gênica de IL- 1ß e TNFα. Microcápsulas implantadas i.p. em camundongos foram recuperadas após 7 ou 30 dias, para análises de biocompatibilidade. A expressão de níveis de mRNA (bax, bad, bcl-2, bcl-XL, xiap, caspase 3, mcp1/ccl2, hsp70, ldh, insulina 1 e 2), proteínas (Bax, Bcl-XL e Xiap) e a atividade de Caspase3 foram avaliadas em ilhotas microencapsuladas com PFC- e LN-Biodritina, após cultura de 48h em condições de normóxia e hipóxia (<2% O2). Camundongos diabéticos foram transplantados com ilhotas encapsuladas nas diferentes formulações e os animais foram monitorados pelas variações de massa corporal, glicêmicas e pela funcionalidade do enxerto (TOTGs). As ilhotas foram recuperadas de animais normo ou hiperglicêmicos e uma análise de biocompatibilidade das cápsulas foi realizada, assim como a avaliação funcional das células-ß. Após o explante, a glicemia dos animais normoglicêmicos foi monitorada para se atestar a eficiência das ilhotas transplantadas. Resultados Microcápsulas de PFC- e LN-Biodritina são tão estáveis e biocompatíveis quanto as de Biodritina. Para ilhotas encapsuladas em ambos os materiais, em normóxia ou hipóxia, observou-se uma modulação gênica que sugere proteção contra apoptose. Adicionalmente, encontrou-se uma diminuição na expressão de genes indicadores de estresse (mcp1, hsp70). Uma diminuição nos níveis de mRNA de ldh foi vista para PFC-Biodritina, mas o oposto foi encontrado para LN-Biodritina. As diferenças encontradas na expressão proteica sugerem o mesmo padrão anti-apoptótico. Caspase3 não foi modulada por nenhum biomaterial. Nos experimentos de transplante, apenas LN-Biodritina levou reversão prolongada do diabetes, com 60% dos animais normoglicêmicos, 198 dias pós-cirurgia, comparado a 9% do grupo Biodritina. O TOTG demonstrou que camundongos transplantados com ilhotas encapsuladas secretaram mais insulina do que controles, 60 (LN-Biodritina) ou 100 (PFC- e LN-Biodritina) dias pós-cirurgia. O explante restabeleceu a hiperglicemia nos camundongos. Microcápsulas recuperadas de animais hiperglicêmicos apresentavam uma extensa adesão celular. Testes de secreção de insulina in vitro demonstraram que somente ilhotas do grupo normoglicêmico responderam às variações da concentração de glicose. Conclusão A adição de moléculas bioativas à Biodritina é capaz de diminuir o estresse em ilhotas isoladas e tem o potencial de melhorar a terapia pelo transplante de ilhotas

Transplantation of microencapsulated islets represents an attractive therapeutical approach to treat type 1 Diabetes Mellitus, accounting for an improved glycemic control and the abolishment of immunosuppressive therapies. However, maintenance of long-term ß-cell viability remains a major problem. During islet isolation, the loss of extracellular matrix interactions and the hypoxic conditions thereafter dramatically affect ß-cell survival and function. Objective To lessen the burden of islet stress and achieve a better outcome in islet transplantation we tested the addition of perfluorocarbon (PFC) or laminin (LN), molecules associated respectively with oxygenation and cell-cell interaction, to Biodritin, an alginate-based material suitable for cell microencapsulation. Methodology To test the stability of PFC-Biodritin and LN-Biodritin composites, microcapsules were subjected to different stresses (rotational, osmotic, temperature and culture) for 7 and 30 days. To assess biomaterial purity microcapsules were co-incubated with RAW264.7 murine macrophage cell line for 3, 9 and 24h and macrophage activation was detected through mRNA levels of IL-1ß and TNFα. Microcapsules were implanted i.p. in mice and retrieved after 7 or 30 days, for biocompatibility analyses. Gene expression at mRNA (bax, bad, bcl-2, bcl-XL, xiap, caspase 3, mcp1/ccl2, hsp70, ldh, insulin 1 and 2) and protein (Bax, Bcl-XL and Xiap) levels, together with Caspase3 activity, were evaluated in islets microencapsulated in PFC- or LN-Biodritin, upon culturing for 48h in normoxic or hypoxic (<2% O2) conditions. Diabetic mice were transplanted with PFC- or LN-Biodritin microencapsulated islets, followed by assessments of body weight, glycemia and graft function by oral glucose tolerance tests (OGTTs). Microencapsulated islets were retrieved from normoglycemic or hyperglycemic mice and biocompatibility analyses of the beads together with a functional assessment of the graft followed. After graft removal, normoglycemic animals had their glycemias monitored to attest the efficacy of the transplanted islets. Results PFC- and LN-Biodritin microcapsules were as stable and biocompatible as Biodritin. For both biomaterials in normoxia and hypoxia a modulation in gene expression was observed in islets associated with a protection against apoptosis. Also, a decreased expression of stress-related genes (mcp1, hsp70) was evidenced. ldh mRNA levels were down-regulated in PFC-Biodritin microencapsulated islets but upregulated in the presence of LN. Increased levels of insulin mRNA were observed. The differences seen in protein expression indicated the same anti-apoptotic pattern. Caspase3 activity was not different between groups. Concerning diabetes reversal experiments, only mice transplanted with LN-Biodritin microencapsulated islets presented a better outcome, with 60% remaining euglycemic at 198 days post-surgery, compared with 9% for the Biodritin group. OGTT showed that mice transplanted with encapsulated islets secreted more insulin than normal mice, 60 (LN-Biodritin) or 100 days (PFC- and LN-Biodritina) posttransplant. Hyperglycemia was achieved after the retrieval of microcapsules showing graft efficacy. Retrieved microcapsules revealed an extensive overgrowth in most beads from hyperglycemic mice. A static glucose stimulated insulin secretion test revealed that only islets from normoglycemic subjects were able to secrete insulin according to glucose concentration. Conclusion- The addition of bioactive molecules to Biodritin may lessen the stress of isolated islets and have the potential to improve islet transplantation therapy.

Comparing the osteogenic potential of canine mesenchymal stem cells derived from adipose tissues, bone marrow, umbilical cord blood, and Wharton's jelly for treating bone defects

Alternative sources of mesenchymal stem cells (MSCs) for replacing bone marrow (BM) have been extensively investigated in the field of bone tissue engineering. The purpose of this study was to compare the osteogenic potential of canine MSCs derived from adipose tissue (AT), BM, umbilical cord blood (UCB), and Wharton's jelly (WJ) using in vitro culture techniques and in vivo orthotopic implantation assays. After canine MSCs were isolated from various tissues, the proliferation and osteogenic potential along with vascular endothelial growth factor (VEGF) production were measured and compared in vitro. For the in vivo assay, MSCs derived from each type of tissue were mixed with beta-tricalcium phosphate and implanted into segmental bone defects in dogs. Among the different types of MSCs, AT-MSCs had a higher proliferation potential and BM-MSCs produced the most VEGF. AT-MSCs and UCB-MSCs showed greater in vitro osteogenic potential compared to the other cells. Radiographic and histological analyses showed that all tested MSCs had similar osteogenic capacities, and the level of new bone formation was much higher with implants containing MSCs than cell-free implants. These results indicate that AT-MSCs, UCB-MSCs, and WJ-MSCs can potentially be used in place of BM-MSCs for clinical bone engineering procedures.

Avaliação radiográfica, biomecânica e microscópica de enxerto ósseo xenogênico associado a implantes de titânio em mandíbula de coelhos / Radiographic, biomechanical and microscopic evaluation of xenogenic bone graft associated to titanium implants in rabbits mandible

O propósito deste trabalho foi avaliar a interação de matriz óssea bovina inorgânica, (Gen-ox®) como material de enxerto e implantes de titânio em mandíbula de coelhos. A amostra constituiu-se de 32 coelhos da raça Botucatu de aproximadamente 4kg. Os incisivos inferiores destes animais foram extraídos e foi realizada um defeito de aproximadamente 4mm de diâmetro. Um dos lados foi mantido apenas com coágulo (grupo controle) e o outro lado (grupo experimental) recebeu o enxerto do material proposto. Após 60 dias foram instalados implantes bilateralmente na região do defeito. Os animais foram eutanaziados imediatamente após a instalação dos implantes e após 1, 2 e 6 meses (n=6) para o grupo microscópico a após 2 e 6 meses para o grupo do teste biomecânico (n=4). A análise constituiu de medidas da distância da crista óssea alveolar à plataforma do implante, da densidade radiográfica em valores de pixel ma região do alvéolo, de teste biomecânico de torque de remoção e microscópico através de medidas de contato osso-implante (BIC) (em µm e %) e área de osso (em µm² e %). Os resultados demonstraram que não houve diferenças significantes nem durante o tempo e nem entre os grupos na analise de distancia entre crista óssea alveolar e plataforma do implante. Na medida de densidade óssea, não houve variação significante ao longo do tempo, mas houve diferença estatisticamente significante entre os grupos, sendo maior no grupo experimental. Para o teste biomecânico também não houve variação significante do torque de remoção ao longo do tempo e nem houve diferença significante entre os grupos. Na análise de contato osso implante (BIC) não houve variação significante ao longo do tempo, mas houve diferença estatisticamente significante entre os ...

The aim of this study was to evaluate the interaction of an inorganic bovine bone matrix (Gen-ox®) as bone graft and titanium implants in rabbits mandible. Thirty-two Botucatu rabbits weighting about 4kg were used on this experiment. The mandible incisives were extracted and a defect of 4 mm of diameter was performed. One socket and defect was filled with experimental material. The opposite site was left to heal naturally and served as control. After 60 days, the implants were installed on the defect region. The animals were killed immediately after the implant installation and after 1, 2 and 6 months. Vertical bone height, bone density, biomechanical test, bone implant contact (BIC) and bone area near to implant threads were evaluated. The results showed that there was no significant differences on the distance between the alveolar bone crest to the implant platform (vertical bone height) along the time, neither between groups. On the bone density (pixel value) there was no significant difference along the time, but there was significant difference between groups, being the experimental group higher. For the biomechanical test there was no significant difference along the time, neither between groups. There was no significant differences on the bone implant contact (BIC) measures along the time, but there was significant difference between groups, being the experimental groups higher and there was interaction between time and groups. On bone area evaluation, there was no significant difference along the time, neither between the groups during the analyzed time. The graft behavior observed by the authors was similar to that of the control group, suggesting high acceptance of the material as graft option associated to titanium implants.

Optical density of bone repair after implantation of homogenous demineralized dentin matrix in diabetic rabbits

This research evaluated the bone repair process after implantation of homogenous demineralized dentin matrix (HDDM) in surgical defects in the parietal bone of rabbits with alloxan-induced diabetes, using a polytetrafluorethylene (PTFe) barrier for guided bone regeneration. Thirty-six rabbits were used and divided into four groups: control (C, n = 12), diabetic (D, n = 12, left parietal bone), diabetic with PTFe (D-PTFe, same 12 rabbits, right parietal bone), and diabetic with PTFe associated to HDDM (D-PTFe+HDDM, n = 12). Bone defects were created in the parietal bone of the rabbits and the experimental treatments were performed, where applicable. The rabbits were sacrificed after 15, 30, 60 and 90 days. The bone defects were examined radiographically and by optical density (ANOVA and Tukey test, p < .05). The radiographic findings showed that the D-PTFe+HDDM group presented greater radiopacity and better trabecular bone arrangement when compared to that of the C, D and D-PTFe groups. The statistical analysis showed significant differences in the optical density of the newly formed bone among the studied groups. It was possible to conclude that HDDM was biocompatible in diabetic rabbits.

Implantation of canine umbilical cord blood-derived mesenchymal stem cells mixed with beta-tricalcium phosphate enhances osteogenesis in bone defect model dogs

This study was performed to evaluate the osteogenic effect of allogenic canine umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) mixed with beta-tricalcium phosphate (beta-TCP) in orthotopic implantation. Seven hundred milligrams of beta-TCP mixed with 1 x 10(6) UCB-MSCs diluted with 0.5 ml of saline (group CM) and mixed with the same volume of saline as control (group C) were implanted into a 1.5 cm diaphyseal defect and wrapped with PLGC membrane in the radius of Beagle dogs. Radiographs of the antebrachium were made after surgery. The implants were harvested 12 weeks after implantation and specimens were stained with H&E, toluidine blue and Villanueva-Goldner stains for histological examination and histomorphometric analysis of new bone formation. Additionally, UCB-MSCs were applied to a dog with non-union fracture. Radiographically, continuity between implant and host bone was evident at only one of six interfaces in group C by 12 weeks, but in three of six interfaces in group CM. Radiolucency was found only near the bone end in group C at 12 weeks after implantation, but in the entire graft in group CM. Histologically, bone formation was observed around beta-TCP in longitudinal sections of implant in both groups. Histomorphometric analysis revealed significantly increased new bone formation in group CM at 12 weeks after implantation (p < 0.05). When applied to the non-union fracture, fracture healing was identified by 6 weeks after injection of UCB-MSCs. The present study indicates that a mixture of UCB-MSCs and beta-TCP is a promising osteogenic material for repairing bone defects.

Imaging Findings of Implanted Absorbable Mesh in Patients with Breast Partial Resection

PURPOSE: The author presents imaging findings of patients that underwent partial resection of the breast followed by absorbable mesh implantation. MATERIALS AND METHODS: Ultrasonographic (n=18) and mammographic (n=11) images of patients that had undergone absorbable mesh implantation after breast partial resection were reviewed retrospectively. Sequential changes of the lesions were analyzed in follow-up ultrasonographic examinations, focusing on the change of the size and pattern of the lesion. The presence of a mass, asymmetry, focal asymmetry, architectural distortion, and calcification were evaluated by mammography. Pathologic findings of the implanted mesh in available cases were analyzed. RESULTS: Ultrasonograms revealed a well-encapsulated anechoic lesion with (pattern 1, n=11) or without (pattern 2, n=5) internal isoechoic nodular portion, and a hyperechoic mass-like lesion without anechoic portion (pattern 3, n=2). The mean length of the longest diameter decreased gradually as determined in follow-up examinations (3 months, 6.12 +/- 2.599cm; 6 months, 5.08 +/- 2.105cm; 12 months, 3.26 +/- 2.206cm). In mammograms, a mass (n=4) was noted at the surgical site and focal asymmetry, overlapping with the postoperative change, was seen in the remaining seven cases. Pathologic findings of two cases revealed foreign body reaction. CONCLUSION: Ultrasonography of the patients that underwent breast partial resection followed by absorbable mesh implantation showed a well-encapsulated cyst at the surgical site that gradually decreased in follow-up examinations. Adjunctive ultrasonography combined with mammography would be recommended in postoperative follow-up examinations.

Avaliaçäo da biocompatibilidade de implantes de Hapset (hidroxiapatita associada ao sulfato de cálcio) em feridas de extraçäo dental. Estudo histológico em ratos / Evalution of the Hapset implants (hidroxyapatie/calcium sulphate) biocompatibility in dental extraction wounds. Histologycal study in rats

A biocompatibilidade de implantes Hapset R (hidroxiapatita associada ao sulfato de cálcio) foi estudada em feridas de extraçäo dental pela inserçäo de tubos de polietileno em alvéolos de ratos após extraçöes, contendo em suas extremidades Hapset e Gutapercha. Os animais foram sacrificados em períodos de 7, 15, 21 e 30 dias após as cirurgias, sendo os espécimes obtidos processados e submetidos a análise histológica. Verificou-se reaçäo mais severa na regiäo contígua ao implante de Hapset, indicando ser este material mais irritante que a Gutapercha. A metodologia empregada permitiu boa análise comparativa entre os materiais empregados, podendo ser empregada em estudos de comportamento biológico de materiais odontológicos

Estudio "in vivo" de los fenómenos de interfase de las prótesis porosas de cadera / Study in living of the interface phenomenon of the hip prothesis of both smooth and porous surface

Fragmentos irregulares de prótesis totales de cadera de superficie lisa y de superficie porosa de última generación protésica tanto comentados como no cementados fueron colocados en diáfisis femoral de perros con períodos de permanencia en los mismos que variaron desde trece (13) días hasta un (1) año, cincuenta y dos (52) días y luego extraídos y examinados bajo microscopia electrónica de barrido a objeto de observar crecimiento de mamelones óseo alejado del tercio proxima del fémur

Biomateriais na substituiçäo de tecidos moles / Biomaterials in soft tissue substitution

Os autores apresentam revisäo da literatura sobre o uso de biomateriais na reparaçäo de tecidos moles. Säo descritos os principais aspectos relacionados ao silicone, poliamida, polietileno e politetrafluoretileno, quando utilizados como inclusöes. Destaques säo dados à resposta inflamatória local, formaçäo de cápsula, fenômenos de sensibilizaçäo. Säo ainda salientadas a interferência das propriedades físico-químicas e superfície na evocaçäo de resposta inflamatória, além da fixaçäo das inclusöes em relaçäo à porosidade das mesmas