scholarly journals Stem Cell Delivery with Polymer Hydrogel for Treatment of Intervertebral Disc Degeneration: From 3D Culture to Design of the Delivery Device for Minimally Invasive Therapy

2016 ◽  
Vol 25 (12) ◽  
pp. 2213-2220 ◽  
Author(s):  
Deepak Kumar ◽  
Alex Lyness ◽  
Irini Gerges ◽  
Christina Lenardi ◽  
Nicholas R. Forsyth ◽  
...  
Keyword(s):  
Minimally Invasive ◽  
Disc Degeneration ◽  
Mechanical Strain ◽  
3D Culture ◽  
Human Mesenchymal Stem Cells ◽  
Intervertebral Discs ◽  
Delivery Device ◽  
Polymer Hydrogel ◽  
Uv Curable ◽  
Stem Cell Delivery

Nucleus pulposus (NP) tissue damage can induce detrimental mechanical strain on the biomechanical performance of intervertebral discs (IVDs), causing subsequent disc degeneration. A novel, photocurable, injectable, synthetic polymer hydrogel (pHEMA-co-APMA grafted with PAA) has already demonstrated success in encapsulating and differentiating human mesenchymal stem cells (hMSCs) toward an NP phenotype during hypoxic conditions. After demonstration of promising results in our previous work, in this study we have further investigated the inclusion of mechanical stimulation and its impact on hMSC differentiation toward an NP phenotype through the characterization of matrix markers such as SOX-9, aggrecan, and collagen II. Furthermore, investigations were undertaken in order to approximate delivery parameters for an injection delivery device, which could be used to transport hMSCs suspended in hydrogel into the IVD. hMSC-laden hydrogel solutions were injected through various needle gauge sizes in order to determine its impact on postinjection cell viability and IVD tissue penetration. Interpretation of these data informed the design of a potential minimally invasive injection device, which could successfully inject hMSCs encapsulated in a UV-curable polymer into NP, prior to photo-cross-linking in situ.

scholarly journals Identification of Novel FNIN2 and FNIN3 Fibronectin-Derived Peptides That Promote Cell Adhesion, Proliferation and Differentiation in Primary Cells and Stem Cells

2021 ◽  
Vol 22 (6) ◽  
pp. 3042
Author(s):  
Eun Ju Lee ◽  
Khurshid Ahmad ◽  
Shiva Pathak ◽  
SunJu Lee ◽  
Mohammad Hassan Baig ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Culture ◽  
Cell Adhesion ◽  
3D Culture ◽  
Human Mesenchymal Stem Cells ◽  
Cell Types ◽  
Primary Cells ◽  
Cell Culture Techniques ◽  
Culture Techniques ◽  
Promote Cell Adhesion

In recent years, a major rise in the demand for biotherapeutic drugs has centered on enhancing the quality and efficacy of cell culture and developing new cell culture techniques. Here, we report fibronectin (FN) derived, novel peptides fibronectin-based intergrin binding peptide (FNIN)2 (18-mer) and FNIN3 (20-mer) which promote cell adhesion proliferation, and the differentiation of primary cells and stem cells. FNIN2 and 3 were designed based on the in silico interaction studies between FN and its receptors (integrin α5β1, αvβ3, and αIIbβ3). Analysis of the proliferation of seventeen-cell types showed that the effects of FNINs depend on their concentration and the existence of expressed integrins. Significant rhodamine-labeled FNIN2 fluorescence on the membranes of HeLa, HepG2, A498, and Du145 cells confirmed physical binding. Double coating with FNIN2 or 3 after polymerized dopamine (pDa) or polymerized tannic acid (pTA) precoating increased HBEpIC cell proliferation by 30–40 percent, suggesting FNINs potently affect primary cells. Furthermore, the proliferation of C2C12 myoblasts and human mesenchymal stem cells (MSCs) treated with FNINs was significantly increased in 2D/3D culture. FNINs also promoted MSC differentiation into osteoblasts. The results of this study offer a new approach to the production of core materials (e.g., cell culture medium components, scaffolds) for cell culture.

scholarly journals Minimally invasive debridement and drainage using intraoperative CT-Guide in multilevel spondylodiscitis: a long‐term follow‐up study

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Jianbiao Xu ◽  
Leiming Zhang ◽  
Rongqiang Bu ◽  
Yankang Liu ◽  
Kai-Uwe Lewandrowski ◽  
...  
Keyword(s):  
Minimally Invasive ◽  
Clinical Manifestations ◽  
Operation Time ◽  
Intervertebral Discs ◽  
Intraoperative Ct ◽  
Long Term Follow Up ◽  
Invasive Method ◽  
Vertebral Bodies ◽  
Adjacent Segments

Abstract Background Spondylodiscitis is an unusual infectious disease, which usually originates as a pathogenic infection of intervertebral discs and then spreads to neighboring vertebral bodies. The objective of this study is to evaluate percutaneous debridement and drainage using intraoperative CT-Guide in multilevel spondylodiscitis. Methods From January 2002 to May 2017, 23 patients with multilevel spondylodiscitis were treated with minimally invasive debridement and drainage procedures in our department. The clinical manifestations, evolution, and minimally invasive debridement and drainage treatment of this refractory vertebral infection were investigated. Results Of the enrolled patients, the operation time ranged from 30 minutes to 124 minutes every level with an average of 48 minutes. Intraoperative hemorrhage was minimal. The postoperative follow-up period ranged from 12 months to 6.5 years with an average of 3.7 years. There was no reactivation of infection in the treated vertebral segment during follow-up, but two patients with fungal spinal infection continued to progress by affecting adjacent segments prior to final resolution. According to the classification system of Macnab, one patient had a good outcome at the final follow-up, and the rest were excellent. Conclusions Minimally invasive percutaneous debridement and irrigation using intraoperative CT-Guide is an effective minimally invasive method for the treatment of multilevel spondylodiscitis.

scholarly journals A Hyaluronan and Platelet-Rich Plasma Hydrogel for Mesenchymal Stem Cell Delivery in the Intervertebral Disc: An Organ Culture Study

2021 ◽  
Vol 22 (6) ◽  
pp. 2963
Author(s):  
Fabrizio Russo ◽  
Luca Ambrosio ◽  
Marianna Peroglio ◽  
Wei Guo ◽  
Sebastian Wangler ◽  
...  
Keyword(s):  
Gene Expression ◽  
Intervertebral Disc ◽  
Organ Culture ◽  
Platelet Rich Plasma ◽  
Cell Activity ◽  
Human Mesenchymal Stem Cells ◽  
Cell Phenotype ◽  
Cytokeratin 19 ◽  
Stem Cell Delivery ◽  
Normal Metabolism

The purpose of the present pilot study was to evaluate the effect of a hydrogel composed of hyaluronic acid (HA) and platelet-rich plasma (PRP) as a carrier for human mesenchymal stem cells (hMSCs) for intervertebral disc (IVD) regeneration using a disc organ culture model. HA was mixed with batroxobin (BTX) and PRP to form a hydrogel encapsulating 1 × 106 or 2 × 106 hMSCs. Bovine IVDs were nucleotomized and filled with hMSCs suspended in ~200 μL of the PRP/HA/BTX hydrogel. IVDs collected at day 0 and nucleotomized IVDs with no hMSCs and/or hydrogel alone were used as controls. hMSCs encapsulated in the hydrogel were also cultured in well plates to evaluate the effect of the IVD environment on hMSCs. After 1 week, tissue structure, scaffold integration, hMSC viability and gene expression of matrix and nucleus pulposus (NP) cell markers were assessed. Histological analysis showed a better preservation of the viability of the IVD tissue adjacent to the gel in the presence of hMSCs (~70%) compared to the hydrogel without hMSCs. Furthermore, disc morphology was maintained, and the hydrogel showed signs of integration with the surrounding tissues. At the gene expression level, the hydrogel loaded with hMSCs preserved the normal metabolism of the tissue. The IVD environment promoted hMSC differentiation towards a NP cell phenotype by increasing cytokeratin-19 (KRT19) gene expression. This study demonstrated that the hydrogel composed of HA/PRP/BTX represents a valid carrier for hMSCs being able to maintain a good cell viability while stimulating cell activity and NP marker expression.

scholarly journals Evaluation of apoptotic cell death in normal and chondrodystrophic canine intervertebral discs

2012 ◽  
Vol 2 (1) ◽  
pp. 6 ◽  
Author(s):  
Marie Klauser ◽  
Franck Forterre ◽  
Marcus Doherr ◽  
Andreas Zurbriggen ◽  
David Spreng ◽  
...  
Keyword(s):  
Nucleus Pulposus ◽  
Disc Degeneration ◽  
Annulus Fibrosus ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
Intervertebral Discs ◽  
Discogenic Pain ◽  
Age Related ◽  
Chondroid Metaplasia ◽  
Related Increase

Disc degeneration occurs commonly in dogs. A variety of factors is thought to contribute an inappropriate disc matrix that isolate cells in the disc and lead to apoptosis. Disc herniation with radiculopathy and discogenic pain are the results of the degenerative process. The objective of this prospective study was to determine the extent of apoptosis in intact and herniated intervertebral discs of chondrodystrophic dogs and non-chondrodystrophic dogs. In addition, the nucleus pulposus (NP) was histologically compared between non-chondrodystrophic and chondrodystrophic dogs. Thoracolumbar intervertebral discs and parts of the extruded nucleus pulposus were harvested from 45 dogs. Samples were subsequently stained with haematoxylin-eosin and processed to detect cleaved caspase-3 and poly(ADP-ribose) polymerase. A significant greater degree of apoptosis was observed in herniated NPs of chondrodystrophic dogs compared to non- chondrodystrophic dogs with poly (ADP-ribose) polymerase and cleaved caspase- 3 detection. Within the group of chondrodystrophic dogs, dogs with an intact disc and younger than 6 years showed a significant lower incidence of apoptosis in the NP compared to the herniated NP of chondrodystrophic dogs. The extent of apoptosis in the annulus fibrosus was not different between the intact disc from chondrodystrophic and non- chondrodystrophic dogs. An age-related increase of apoptotic cells in NP and annulus fibrosus was found in the intact non-herniated intervertebral discs. Histologically, absence of notochordal cells and occurrence of chondroid metaplasia were observed in the nucleus pulposus of chondrodystrophic dogs. As a result, we found that apoptosis plays a role in disc degeneration in chondrodystrophic dogs.

scholarly journals Excessive mechanical strain accelerates intervertebral disc degeneration by disrupting intrinsic circadian rhythm

2021 ◽  
Author(s):  
Sheng-Long Ding ◽  
Tai-Wei Zhang ◽  
Qi-Chen Zhang ◽  
Wang Ding ◽  
Ze-Fang Li ◽  
...  
Keyword(s):  
Circadian Rhythm ◽  
Intervertebral Disc ◽  
Mechanical Loading ◽  
Disc Degeneration ◽  
Intervertebral Disc Degeneration ◽  
Night Shift ◽  
Mechanical Strain ◽  
Inhibitory Effect ◽  
Shift Workers ◽  
Normal Environment

AbstractNight shift workers with disordered rhythmic mechanical loading are more prone to intervertebral disc degeneration (IDD). Our results showed that circadian rhythm (CR) was dampened in degenerated and aged NP cells. Long-term environmental CR disruption promoted IDD in rats. Excessive mechanical strain disrupted the CR and inhibited the expression of core clock proteins. The inhibitory effect of mechanical loading on the expression of extracellular matrix genes could be reversed by BMAL1 overexpression in NP cells. The Rho/ROCK pathway was demonstrated to mediate the effect of mechanical stimulation on CR. Prolonged mechanical loading for 12 months affected intrinsic CR genes and induced IDD in a model of upright posture in a normal environment. Unexpectedly, mechanical loading further accelerated the IDD in an Light-Dark (LD) cycle-disrupted environment. These results indicated that intrinsic CR disruption might be a mechanism involved in overloading-induced IDD and a potential drug target for night shift workers.

Differential osteogenicity of multiple donor-derived human mesenchymal stem cells and osteoblasts in monolayer, scaffold-based 3D culture and in vivo

2016 ◽  
Vol 61 (3) ◽  
pp. 253-266 ◽  
Author(s):  
Verena M.C. Quent ◽  
Christina Theodoropoulos ◽  
Dietmar W. Hutmacher ◽  
Johannes C. Reichert
Keyword(s):  
Matrix Protein ◽  
3D Culture ◽  
Human Mesenchymal Stem Cells ◽  
Cell Types ◽  
Matrix Mineralization ◽  
Dentin Matrix Protein ◽  
Cellular Models ◽  
Parathyroid Hormone Related Protein ◽  
Study Results

Abstract We set out to compare the osteogenicity of human mesenchymal stem (hMSCs) and osteoblasts (hOBs). Upon osteogenic induction in monolayer, hMSCs showed superior matrix mineralization expressing characteristic bone-related genes. For scaffold cultures, both cell types presented spindle-shaped, osteoblast-like morphologies forming a dense, interconnected network of high viability. On the scaffolds, hOBs proliferated faster. A general upregulation of parathyroid hormone-related protein (PTHrP), osteoprotegrin (OPG), receptor activator of NF-κB ligand (RANKL), sclerostin (SOST), and dentin matrix protein 1 (DMP1) was observed for both cell types. Simultaneously, PTHrP, RANKL and DMP-1 expression decreased under osteogenic stimulation, while OPG and SOST increased significantly. Following transplantation into NOD/SCID mice, μCT and histology showed increased bone deposition with hOBs. The bone was vascularized, and amounts further increased for both cell types after recombinant human bone morphogenic protein 7 (rhBMP-7) addition also stimulating osteoclastogenesis. Complete bone organogenesis was evidenced by the presence of osteocytes and hematopoietic precursors. Our study results support the asking to develop 3D cellular models closely mimicking the functions of living tissues suitable for in vivo translation.

Formation of Wrinkle Patterns on Porous Elastomeric Membrane and Their Fabrication of Hierarchical Architectures

2008 ◽  
Vol 1129 ◽  
Author(s):  
Yue Cui ◽  
Shu Yang
Keyword(s):  
Cell Attachment ◽  
Mechanical Strain ◽  
Crystal Formation ◽  
Plasma Oxidation ◽  
Oxidation Condition ◽  
Uv Curable ◽  
Replica Molding ◽  
Hierarchical Architectures ◽  
Elastomeric Membrane ◽  
Mechanical Stretching

AbstractWe report the formation of wrinkle patterns on porous elastomeric membrane and their fabrication of hierarchical architectures through mechanical stretching and replica molding. The technique builds upon a buckling instability of a stiff layer supported by a porous elastomeric membrane which was induced by surface plasma oxidation of the pre-stretched porous elastomer followed by removal of the applied mechanical strain to form wrinkle patterns, and replica molding of the deformed features on the porous membrane into epoxy to form hierarchical architectures through casting the UV-curable epoxy prepolymer and UV curing. We find that due to the existence of micropores on the membrane, the formation of wrinkle patterns is different from that formed on a continuous elastomeric film, and by varying the applied mechanical stretching strain condition and plasma oxidation condition, the wrinkle patterns could be either confined by the micropores on the membrane to exhibit a wavelength equal to its pitch or form wrinkles with much large wavelength compared with that formed on a continuous elastomeric film. Therefore, the micropillar arrays fabricated by replica molding could stand on different types of wrinkle patterns to form different hierarchical architectures. The method we illustrate here offers a simple and cost-effective approach to fabricate various hierarchical structures, and provides possibilities for potential applications in various fields, such as microfluidics, micro- and nanofabrication of complex structures, crystal formation, cell attachment, superhydrophobicity and dry adhesion.

scholarly journals Outcomes in cases of lumbar degenerative spondylolisthesis more than 5 years after treatment with minimally invasive decompression: examination of pre- and postoperative slippage, intervertebral disc changes, and clinical results

2016 ◽  
Vol 24 (3) ◽  
pp. 367-374 ◽  
Author(s):  
Gen Mori ◽  
Yasuo Mikami ◽  
Yuji Arai ◽  
Takumi Ikeda ◽  
Masateru Nagae ◽  
...  
Keyword(s):  
Minimally Invasive ◽  
Intervertebral Disc ◽  
Disc Degeneration ◽  
Intervertebral Disc Degeneration ◽  
Degenerative Spondylolisthesis ◽  
Clinical Results ◽  
Natural Course ◽  
Disc Height ◽  
Minimally Invasive Decompression

OBJECT There are reports that fusion is the standard treatment of choice for cases of lumbar degenerative spondylolisthesis (LDS) associated with lumbar spinal canal stenosis with a large degree of slippage. The reasons why, however, have not been clarified. On the other hand, it is known that the progress of slippage decreases and restabilization occurs over the natural course of LDS. Therefore, if minimally invasive decompression could be performed, there would be little possibility of it influencing the natural course of LDS, so it would not be necessary to include preoperative percentage slip in the criteria for the selection of fusion. This study examined the course of LDS cases more than 5 years after treatment with minimally invasive decompression to determine whether pre- and postoperative slippage and disc changes influence the clinical results. METHODS A total of 51 intervertebral segments in 51 cases with the chief complaint of radicular or cauda equina symptoms due to lumbar spinal canal stenosis were examined after prospective treatment with minimally invasive decompression for LDS. The mean age of the patients at the time of surgery was 66.7 years and the mean follow-up period was 7 years 4 months. Minimally invasive decompression was performed regardless of the degree of low-back pain or percentage slip. The outcome variables were clinical results and changes in imaging findings. RESULTS Over the follow-up period, postoperative percentage slip increased and disc height decreased, but the Japanese Orthopaedic Association score improved. Regardless of the preoperative percentage slip, disc height, or degree of intervertebral disc degeneration or segmental instability, the clinical results were favorable. In the high preoperative percentage slip group, low disc height group, and progressive disc degeneration group, there was little postoperative progress of slippage. In the group with a postoperative slippage increase of more than 5%, slippage increased significantly at postoperative year 2, but no significant difference was observed at the final follow-up. CONCLUSIONS When minimally invasive decompression was performed to treat LDS, the postoperative change in slippage was no different from that during the natural course. Furthermore, regardless of the degree of preoperative slippage or intervertebral disc degeneration, the clinical results were favorable. Also, the higher the preoperative percentage slip and the more that disc degeneration progressed, the more the progress of postoperative slippage decreased. Because the postoperative progress of slippage decreased, it is believed that even after minimally invasive decompression, restabilization occurs as it would during the natural course. If minimally invasive decompression can be performed to treat LDS, it is believed that preoperative percentage slip and intervertebral disc degeneration do not have to be included in the appropriateness criteria for fusion.

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