Bioinspired mineralization of a functionalized injectable dense collagen hydrogel through silk sericin incorporation

2019 ◽  
Vol 7 (3) ◽  
pp. 1064-1077 ◽  
Author(s):  
Gabriele Griffanti ◽  
Wenge Jiang ◽  
Showan N. Nazhat

The incorporation of silk sericin into injectable dense collagen hydrogels represents a powerful approach to mimic the biomineralization process, together with the osteogenic stimulation of seeded mesenchymal stem cells, in vitro.

2019 ◽  
Vol 12 (6) ◽  
pp. 273-279 ◽  
Author(s):  
Jirun Apinun ◽  
Sittisak Honsawek ◽  
Somsak Kuptniratsaikul ◽  
Jutarat Jamkratoke ◽  
Sorada Kanokpanont

AbstractBackgroundSilk fibroin (SF) can be processed into a hydrogel. SF/collagen hydrogel may be a suitable biomaterial for bone tissue engineering.ObjectivesTo investigate in vitro biocompatibility and osteogenic potential of encapsulated rat bone marrow-derived mesenchymal stem cells (rat MSCs) in an injectable Thai SF/collagen hydrogel induced by oleic acid–poloxamer 188 surfactant mixture in an in vitro pilot study.MethodsRat MSCs were encapsulated in 3 groups of hydrogel scaffolds (SF, SF with 0.05% collagen [SF/0.05C], and SF with 0.1% collagen [SF/0.1C]) and cultured in a growth medium and an osteogenic induction medium. DNA, alkaline phosphatase (ALP) activity, and calcium were assayed at periodically for up to 5 weeks. After 6 weeks of culture the cells were analyzed by scanning electron microscopy and energy dispersive spectroscopy.ResultsAlthough SF hydrogel with collagen seems to have less efficiency to encapsulate rat MSCs, their plateau phase growth in all hydrogels was comparable. Inability to maintain cell viability as cell populations declined over 1–5 days was observed. Cell numbers then plateaued and were maintained until day 14 of culture. ALP activity and calcium content of rat MSCs in SF/collagen hydrogels were highest at day 21. An enhancing effect of collagen combined with the hydrogel was observed for proliferation and matrix formation; however, benefits of the combination on osteogenic differentiation and biomineralization are as yet unclear.ConclusionRat MSCs in SF and SF/collagen hydrogels showed osteogenic differentiation. Accordingly, these hydrogels may serve as promising scaffolds for bone tissue engineering.


2020 ◽  
Vol 8 (21) ◽  
pp. 4680-4693
Author(s):  
Jirong Yang ◽  
Yumei Xiao ◽  
Zizhao Tang ◽  
Zhaocong Luo ◽  
Dongxiao Li ◽  
...  

The different negatively charged microenvironments of collagen hydrogels affect the protein adsorption, cell morphology, and chondrogenic differentiation of BMSCs in vitro and in vivo.


2015 ◽  
Vol 137 (1) ◽  
Author(s):  
T. J. Vaughan ◽  
M. Voisin ◽  
G. L. Niebur ◽  
L. M. McNamara

Mechanical loading directs the differentiation of mesenchymal stem cells (MSCs) in vitro and it has been hypothesized that the mechanical environment plays a role in directing the cellular fate of MSCs in vivo. However, the complex multicellular composition of trabecular bone marrow means that the precise nature of mechanical stimulation that MSCs experience in their native environment is not fully understood. In this study, we developed a multiscale model that discretely represents the cellular constituents of trabecular bone marrow and applied this model to characterize mechanical stimulation of MCSs in vivo. We predicted that cell-level strains in certain locations of the trabecular marrow microenvironment were greater in magnitude (maximum ε12 = ∼24,000 με) than levels that have been found to result in osteogenic differentiation of MSCs in vitro (>8000 με), which may indicate that the native mechanical environment of MSCs could direct cellular fate in vivo. The results also showed that cell–cell adhesions could play an important role in mediating mechanical stimulation within the MSC population in vivo. The model was applied to investigate how changes that occur during osteoporosis affected mechanical stimulation in the cellular microenvironment of trabecular bone marrow. Specifically, a reduced bone volume (BV) resulted in an overall increase in bone deformation, leading to greater cell-level mechanical stimulation in trabecular bone marrow (maximum ε12 = ∼48,000 με). An increased marrow adipocyte content resulted in slightly lower levels of stimulation within the adjacent cell population due to a shielding effect caused by the more compliant behavior of adipocytes (maximum ε12 = ∼41,000 με). Despite this reduction, stimulation levels in trabecular bone marrow during osteoporosis remained much higher than those predicted to occur under healthy conditions. It was found that compensatory mechanobiological responses that occur during osteoporosis, such as increased trabecular stiffness and axial alignment of trabeculae, would be effective in returning MSC stimulation in trabecular marrow to normal levels. These results have provided novel insight into the mechanical stimulation of the trabecular marrow MSC population in both healthy and osteoporotic bone, and could inform the design three-dimensional (3D) in vitro bioreactor strategies techniques, which seek to emulate physiological conditions.


2015 ◽  
Vol 18 (1) ◽  
pp. 3-11 ◽  
Author(s):  
J. Nicpoń ◽  
A. Siudzińska ◽  
M. Marędziak ◽  
A. Śmieszek ◽  
K. Basińska ◽  
...  

Abstract The influences of NSAIDs (Nonsteroidal Anti-inflammatory Drugs) – non-selective metamizole and selectively-acting tolfenamic acid were estimated on morphology, ultrastructure, and cytophysiological activity of canine (Ca) and equine (Eq) adipose-derived mesenchymal stem cells (ASCs). The lowest concentration of metamizole (0.01 mg/mL) stimulated the viability and cytophysiological activity of Ca ASCs and did not affect cell morphology. Stimulated cells possessed a proper, fibroblastic shape, with large, eccentrically located nuclei. Similar effects to those observed in Ca ASCs were found in Eq cells treated with both drugs. Cells cultivated with the intermediate (0.1 mg/mL) doses of NSAIDs displayed proper cell morphology, whereas cells cultivated in intermediate dose (0.1 mg/mL) became more flattened. The highest concentrations (1 mg/mL) of both drugs resulted in a cytotoxic effect in Ca and Eq ASCs. Based on these results, we conclude that stimulation of Ca and Eq ASCs with metamizole as well as Eq ASCs with tolfenamic acid can lead to positive effects only when the lowest drug concentrations are applied. This study indicates a different cellular response of canine and equine ASCs treated with metamizole and tolfenamic acid. The obtained data might be potentially useful in the study of functionalized veterinary biomaterials.


2018 ◽  
Vol 2018 ◽  
pp. 1-12 ◽  
Author(s):  
Rosa L. Garnica-Gutiérrez ◽  
Luis A. Lara-Martínez ◽  
Eduardo Palacios ◽  
Felipe Masso ◽  
Alejandra Contreras ◽  
...  

The effects of acid-functionalized and polycitric acid- (PCA-) polymerized carbon nanotubes (CNTs) in contact with the extracellular membrane of mesenchymal stem cells (MSC), a genetically unmodified cell line with differentiation capability, was evaluated with different cellular parameters. The modified CNTs show differences in the analyzed biological behaviors, that is, intracellular incorporation, cell proliferation, apoptosis, and cytotoxicity as compared with those unpolymerized nanotubes. Due to the reduced cellular uptake of polymerized CNTs, PCA-polymerized CNTs are less cytotoxic and are associated with less apoptotic cell death than the acid-functionalized ones. The effects of nitrogen-doped CNTs (CNx) is also reported, showing that functionalized undoped CNTs present strong stimulation of cell proliferation, whereas functionalized and polymerized CNxs stimulate an apoptotic behavior. The study of MSCs in contact with CNTs and PCA is challenging due to the complexity of its various signaling components. Our results provide basis for further studies aimed to understand the relevant role that the interaction of these nanotubes with extracellular membrane could have a crucial structure for tissue grafting.


2019 ◽  
Vol 98 (9) ◽  
pp. 350-355

Introduction: There is evidence that mesenchymal stem cells (MSCs) could trans-differentiate into the liver cells in vitro and in vivo and thus may be used as an unfailing source for stem cell therapy of liver disease. Combination of MSCs (with or without their differentiation in vitro) and minimally invasive procedures as laparoscopy or Natural Orifice Transluminal Endoscopic Surgery (NOTES) represents a chance for many patients waiting for liver transplantation in vain. Methods: Over 30 millions of autologous MSCs at passage 3 were transplanted via the portal vein in an eight months old miniature pig. The deposition of transplanted cells in liver parenchyma was evaluated histologically and the trans-differential potential of CM-DiI labeled cells was assessed by expression of pig albumin using immunofluorescence. Results: Three weeks after transplantation we detected the labeled cells (solitary, small clusters) in all 10 samples (2 samples from each lobe) but no diffuse distribution in the samples. The localization of CM-DiI+ cells was predominantly observed around the portal triads. We also detected the localization of albumin signal in CM-DiI labeled cells. Conclusion: The study results showed that the autologous MSCs (without additional hepatic differentiation in vitro) transplantation through the portal vein led to successful infiltration of intact miniature pig liver parenchyma with detectable in vivo trans-differentiation. NOTES as well as other newly developed surgical approaches in combination with cell therapy seem to be very promising for the treatment of hepatic diseases in near future.


2013 ◽  
Author(s):  
Melo Ocarino Natalia de ◽  
Silvia Silva Santos ◽  
Lorena Rocha ◽  
Juneo Freitas ◽  
Reis Amanda Maria Sena ◽  
...  

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