Self-assembled cell sheets composed of mesenchymal stem cells and gelatin nanofibers for the treatment of full-thickness wounds

2020 ◽  
Vol 8 (16) ◽  
pp. 4535-4544
Author(s):  
Oanh-Vu Pham-Nguyen ◽  
Ji Un Shin ◽  
Hyesung Kim ◽  
Hyuk Sang Yoo
Keyword(s):  
Stem Cells ◽  
Wound Healing ◽  
Mesenchymal Stem Cells ◽  
Cell Sheet ◽  
Full Thickness ◽  
Cell Sheets ◽  
Self Assembled

Gelatin-layered PCL nanofibrils for 3D cell sheet formation were composed with adipocyte-derived stem cells for wound healing.

scholarly journals The Combined Thermoresponsive Cell-Imprinted Substrate, Induced Differentiation, and "KLC Sheet" Formation

2021 ◽  
Author(s):  
Neda Keyhanvar ◽  
Nosratollah Zarghami ◽  
Alexander Seifalian ◽  
Peyman Keyhanvar ◽  
Rana Sarvari ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Cell Sheet ◽  
Pdms Substrate ◽  
Cell Sheets ◽  
Dimethyl Siloxane ◽  
Environment Temperature ◽  
Plastic Surface ◽  
Keratinocyte Cell ◽  
Cytokeratin 14

Purpose: Stem cells can exhibit restorative effects with the commitment to functional cells. Cell-imprinted topographies provide adaptable templates and certain dimensions for the differentiation and bioactivity of stem cells. Cell sheet technology using the thermo-responsive polymers detaches the "cell sheets" easier with less destructive effects on the extracellular matrix (ECM). Here, we aim to dictate keratinocyte-like differentiation of mesenchymal stem cells by using combined cell imprinting and sheet technology. Methods: We developed the poly dimethyl siloxane (PDMS) substrate having keratinocyte cell-imprinted topography grafted with the PNIPAAm polymer. Adipose tissue-derived mesenchymal stem cells (AT-MSCs) were cultured on PDMS substrate for 14 days and keratinocyte-like differentiation monitored via the expression of involucrin, P63, and cytokeratin 14. Results: Data showed the efficiency of the current protocol in the fabrication of PDMS molds. The culture of AT-MSCs induced typical keratinocyte morphology and up-regulated the expression of cytokeratin-14, Involucrin, and P63 compared to AT-MSCs cultured on the plastic surface (p<0.05). Besides, KLC sheets were generated once slight changes occur in the environment temperature. Conclusion: These data showed the hypothesis that keratinocyte cell imprinted substrate can orient AT-MSCs toward KLCs by providing a specific niche and topography.

Studies on the Efficacy of Single and Twice Application of Mesenchymal Stem Cells in Full Thickness Cutaneous Wound Healing

2021 ◽  
Author(s):  
Jyotsana Bhatt ◽  
Amarpal . ◽  
Abas Rashid Bhat ◽  
Anuj Pratap Singh ◽  
A.M. Pawde ◽  
...  
Keyword(s):  
Stem Cells ◽  
Wound Healing ◽  
Mesenchymal Stem Cells ◽  
Skin Wound ◽  
Full Thickness ◽  
Cutaneous Wound ◽  
Group Iii ◽  
Group I ◽  
Thickness Skin ◽  
Early Wound

Background: A lot of research has been done in the field of wound healing utilising stem cell therapy. This study was conducted to compare the efficacy of single and twice application of mesenchymal stem cells for cutaneous wound healing.Methods: The present study was done on 18 guinea pigs and a 2.5 X 2.5 cm2 full-thickness skin wound was created on the dorsum of each under standard anesthetic protocol. Animals were divided into 3 groups having 6 animals in each. The animals of group I were administered phosphate buffer saline (PBS) subcutaneously on day 0, group II received 1×106 mesenchymal stem cells (MSCs) in PBS subcutaneously on day 0 and group III received 1×106 MSCs in PBS subcutaneously on day 0 and again at day 3, at wound margins. Clinical, photographic, histopathological and histochemical parameters were recorded for each animal.Result: An overall gross and histopathological evaluation suggested early wound closer, better tissue granulation, early orientation of fibrocytes and collagen fibers, revealing a superior quality healing in animals of group III as compared to rest of the groups. From the above study it was concluded that twice application of MSCs leads to faster and qualitatively better healing. 

scholarly journals Experimental evaluation of the effectiveness of adipose mesenchymal stem cells in full-thickness skin wounds

2021 ◽  
Vol 9 (2) ◽  
pp. 175-181
Author(s):  
Ilya V. Artsimovich ◽  
Evgenii V. Zinoviev ◽  
Aleksander V. Gostimskii ◽  
Marat S. Asadulaev ◽  
Sergey V. Vissarionov ◽  
...  
Keyword(s):  
Stem Cells ◽  
Wound Healing ◽  
Mesenchymal Stem Cells ◽  
Control Group ◽  
Skin Defect ◽  
Full Thickness ◽  
Skin Defects ◽  
Full Thickness Skin ◽  
Healing Index ◽  
Thickness Skin

BACKGROUND: Currently, numerous techniques or medical devices that allow complete or partial restoration of the lost skin within a short time remain as subjects of development. Many studies have shown the effectiveness of using stem cells in the treatment of full-thickness skin defects, but their use remains very limited. At present, there is no consensus among researchers about the advisability of the use of stem cells in the treatment of burns as well as about the method of their introduction. AIM: This study aimed to examine the possibility of accelerating the reparative histogenesis of tissues in the zone of deep skin burns using cultures of adipogenic mesenchymal cells, as well as to evaluate the effectiveness of various methods of introducing cultures of these cells into the defect zone. MATERIALS AND METHODS: An experimental study was carried out on small laboratory animals (rats). After simulating a grade III burn, mesenchymal stem cells were transplanted and superficially applied to the wound surface or injected under the fascia. In the control group, no wound treatment was performed. To assess the effectiveness of the wound-healing preparations, the appearance of wounds was assessed daily, the nature of the discharge and presence and type of granulations were noted, and the timing of scab rejection and wound healing was recorded. The wound area was assessed using a planimetric method. A histological examination of wound biopsies was carried out on days 7, 14, 21, and 28 of observation. RESULTS: The application of adipogenic mesenchymal stem cells demonstrated the greatest efficiency on the developed burn model. Subfascial administration was less effective, but this method achieved a significant acceleration of wound healing in comparison with the control group. An increase in the healing index by 56.6% demonstrates the highest intensity of reparative regeneration in animals applied with adipogenic mesenchymal stem cells. CONCLUSIONS: The preliminary results show that the application of adipogenic mesenchymal stem cells on the skin defect is more effective than subfascial administration based on the healing index. The use of adipogenic mesenchymal stem cells may substantially increase the effectiveness of the treatment of full-thickness skin defects.

scholarly journals Role of Adipose-Derived Mesenchymal Stem Cells in Full Thickness Wound Healing

2017 ◽  
Vol 07 (09) ◽  
Author(s):  
Iswinarno Dososaputro ◽  
Lisette T Hoekstra ◽  
Dinar Rahmania ◽  
David S Perdanakusuma
Keyword(s):  
Stem Cells ◽  
Wound Healing ◽  
Mesenchymal Stem Cells ◽  
Full Thickness ◽  
Full Thickness Wound

scholarly journals Effect of Gelatin on Osteogenic Cell Sheet Formation Using Canine Adipose-Derived Mesenchymal Stem Cells

2017 ◽  
Vol 26 (1) ◽  
pp. 115-123 ◽  
Author(s):  
Ah Young Kim ◽  
Yongsun Kim ◽  
Seung Hoon Lee ◽  
Yongseok Yoon ◽  
Wan-Hee Kim ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Proliferation ◽  
Mesenchymal Stem Cells ◽  
Bone Regeneration ◽  
Cell Sheet ◽  
Proliferation Rate ◽  
Osteogenic Cell ◽  
Cell Sheets ◽  
Gene Markers ◽  
Cell Proliferation Rate

Osteogenically differentiated cell sheet techniques using mesenchymal stem cells (MSCs) are available to stimulate bone regeneration. The advantage of the cell sheet technique is delivering live cells effectively into the focal region. We developed a novel osteogenic cell sheet technique by adding gelatin to osteogenic cell medium. Gelatin-induced osteogenic cell sheets (GCSs) were compared to conventional osteogenic cell sheets (OCSs). Undifferentiated MSCs (UCs) were used as a control. The morphology of these cell sheets was evaluated microscopically and histologically. The time-dependent cell proliferation rate was estimated by DNA quantification. The expression of osteogenic gene markers and the number of calcium depositions were assessed by quantitative real-time polymerase chain reaction and Alizarin red S (ARS) staining, respectively. GCSs were thicker and stronger than OCSs. GCSs showed a significantly higher cell proliferation rate compared to OCSs ( p < 0.05). GCSs exhibited significantly higher upregulation of BMP-7 mRNA compared to OCSs ( p < 0.05). Both GCSs and OCSs showed negative ARS reactivity on day 10, but only GCSs showed positive ARS reactivity on day 21. With this technique, we observed active cell proliferation with abundant ECM and upregulation of osteogenic bone markers, and our results suggest that GCSs could be promising for therapeutic applications in bone regeneration.

scholarly journals Hypoxia Mesenchymal Stem Cells Accelerate Wound Closure Improvement by Controlling α-smooth Muscle actin Expression in the Full-thickness Animal Model

2021 ◽  
Vol 9 (A) ◽  
pp. 35-41
Author(s):  
Nur Fitriani Hamra ◽  
Agung Putra ◽  
Arya Tjipta ◽  
Nur Dina Amalina ◽  
Taufiqurrachman Nasihun
Keyword(s):  
Stem Cells ◽  
Wound Healing ◽  
Mesenchymal Stem Cells ◽  
Smooth Muscle ◽  
Wound Closure ◽  
Smooth Muscle Actin ◽  
Muscle Actin ◽  
Full Thickness ◽  
Treatment Groups ◽  
All Treatment

BACKGROUND: The active myofibroblast producing extracellular matrix deposition regarding wound closure is characterized by alpha-smooth muscle actin (α-SMA) expression. However, the persistence of α-SMA expression due to prolonged inflammation may trigger scar formation. A new strategy to control α-SMA expression in line with wound closure improvement uses hypoxic mesenchymal stem cells (HMSCs) due to their ability to firmly control inflammation for early initiating cell proliferation, including the regulation of α-SMA expression associated with wound closure acceleration. AIM: This study aimed to explore the role of HMSCs in accelerating the optimum wound closure percentages through controlling the α-SMA expression. MATERIALS AND METHODS: Twenty-four full-thickness rats wound model were randomly divided into four groups: Sham (Sh), Control (C) by NaCl administration only, and two treatment groups by HMSCs at doses of 1.5×106 cells (T1) and HMSCs at doses of 3×106 cells (T2). HMSCs were incubated under hypoxic conditions. The α-SMA expression was analyzed under immunohistochemistry staining assay, and the wound closure percentage was analyzed by ImageJ software. RESULTS: This study showed a significant increase in wound closure percentage in all treatment groups that gradually initiated on days 6 and 9 (p < 0.05). In line with the increase of wound closure percentages on day 9, there was also a significant decrease in α-SMA expression in all treatment groups (p < 0.05), indicating the optimum wound healing has preceded. CONCLUSION: HMSCs have a robust ability to accelerated wound closure improvement to the optimum wound healing by controlling α-SMA expression depending on wound healing phases.

scholarly journals Decellularized Articular Cartilage Microparticles for Expansion of Mesenchymal Stem Cells and Zonal Regeneration of Articular Cartilage

2021 ◽  
Author(s):  
Azadeh Sepahvandi ◽  
Safaa Ibrahim Kader ◽  
Victor Anthony Madormo ◽  
Mehri Monavarian ◽  
Esmaiel Jabbari
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Articular Cartilage ◽  
Cell Sheet ◽  
Compressive Modulus ◽  
Human Mscs ◽  
Cell Sheets ◽  
Chondrogenic Medium ◽  
Alginate Gel ◽  
Stratified Structure

Introduction: The objective was to create multilayer cellular constructs using fetal or adult, decellularized articular cartilage in particulate form as microcarriers for expansion and fusion of mesenchymal stem cells (MSCs) to regenerate the stratified structure of articular cartilage. Methods:  Porous microparticles (CMPs) generated from decellularized fetal or adult bovine articular cartilage were used as microcarriers for expansion of human MSCs. The CMP expanded MSCs (CMP-MSCs) were used to generate injectable hydrogels or preformed multilayer constructs for articular cartilage regeneration. In the injectable approach, CMP-MSCs were suspended in alginate gel, crosslinked with calcium chloride, and incubated in chondrogenic medium to generate an injectable regenerative construct. In the preformed approach, fetal or adult CMP-MSCs were suspended in a culture medium, allowed to settle sequentially by the force of gravity, and fused by incubation in chondrogenic medium to generate multilayer cell sheets. The constructs were characterized with respect to compressive modulus, cellularity, and expression of chondrogenic markers. Results: Human MSCs expanded on fetal or adult CMPs in basal medium maintained the expression of mesenchymal markers. The injectable CMP-MSCs hydrogels had significantly higher expression of chondrogenic markers and compressive modulus after four weeks incubation in chondrogenic medium compared to MSCs directly encapsulated in alginate gel; preformed CMP-MSCs cell sheets had significantly higher compressive modulus and expression of chondrogenic markers compared to MSCs in the pellet culture. Conclusion: The preformed cell sheet approach is potentially useful for creating multilayer constructs by sequential gravitational settling of CMP-MSCs to mimic the stratified structure of articular cartilage.

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