Synergistic regenerative therapy of thin endometrium by human placenta-derived mesenchymal stem cells encapsulated within hyaluronic acid hydrogels

Abstract Background: Endometrial injury is one of the major causes of thin endometrium and subfertility. Stem cell-based therapies have made strides towards further efficacious treatment of injured endometrium. However, reported therapeutic stem cells that can be used for thin endometrium are difficult to acquire for large-scale clinical application. The human placenta-derived mesenchymal stem cells (HP-MSCs) are emerging alternative sources of MSCs for their robuster expansion ability, lower immunogenicity as well as extensive sources. To maximize their retention inside the uterus, we loaded HP-MSCs with cross-linked hyaluronic acid hydrogel (HA hydrogel) to investigate their therapeutic efficacy and possible underlying mechanisms.Methods: The murine endometrial injury model was established by ethanol (95%) perfusion, with further intrauterine instillation of treating materials. The retention time of HP-MSCs was detected by in vivo imaging and ex vivo frozen section. Functional restoration of the uterus was assessed by testing embryo implantation rates. The endometrial morphological alteration was observed by H&E staining, Masson staining, and immunohistochemistry (Ki67). The stromal and glandular cells were isolated from the human endometrium to determine proliferation, migration, signaling pathway changes via EdU assay, transwell migration assay, and western blot respectively. Results: Instilled HP-MSCs with HA hydrogel (HP-MSCs-HA) exhibited a prolonged retention time in mouse uteri compared with normal HP-MSCs. In vitro data showed that the HP-MSCs-HA could significantly increase the gland number and endometrial thickness, decrease fibrous area, promote the proliferation of endometrial cells, and improve the embryo implantation rate. In vitro assays indicated that HP-MSCs-HA could not only promote the proliferation and migration of human endometrial stromal via the JNK/Erk1/2-Stat3-VEGF pathway but also promote the proliferation of glandular cells via Jak2-Stat5 and c-Fos-VEGF pathway. Conclusion: Our study suggested the potential therapeutic effects and the underlying mechanisms of HP-MSCs-HA on treating thin endometrium. HA hydrogel could be a preferable delivery method for HP-MSCs and the strategy represents a promising therapeutic approach against endometrial injury in clinical settings.

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Transplantation of umbilical cord-derived mesenchymal stem cells promotes the recovery of thin endometrium in rats

Scientific Reports ◽

10.1038/s41598-021-04454-7 ◽

2022 ◽

Vol 12 (1) ◽

Author(s):

Lu Zhang ◽

Ying Li ◽

Yi-Chao Dong ◽

Chun-Yi Guan ◽

Shi Tian ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Umbilical Cord ◽

Embryo Implantation ◽

Functional Restoration ◽

Model Group ◽

Thin Endometrium ◽

Qrt Pcr ◽

Endometrial Injury ◽

Injury And Repair

AbstractThe endometrium plays a critical role in embryo implantation and pregnancy, and a thin uterus is recognized as a key factor in embryo implantation failure. Umbilical cord mesenchymal stem cells (UC-MSCs) have attracted interest for the repair of intrauterine adhesions. The current study investigated the repair of thin endometrium in rats using the UC-MSCs and the mechanisms involved. Rats were injected with 95% ethanol to establish a model of thin endometrium. The rats were randomly divided into normal, sham, model, and UC-MSCs groups. Endometrial morphological alterations were observed by hematoxylin–eosin staining and Masson staining, and functional restoration was assessed by testing embryo implantation. The interaction between UC-MSCs and rat endometrial stromal cells (ESCs) was evaluated using a transwell 3D model and immunocytochemistry. Microarray mRNA and miRNA platforms were used for miRNA-mRNA expression profiling. Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) analyses were performed to identify the biological processes, molecular functions, cellular components, and pathways of endometrial injury and UC-MSCs transplantation repair and real-time quantitative reverse transcription PCR (qRT-PCR) was performed to further identify the expression changes of key molecules in the pathways. Endometrium thickness, number of glands, and the embryo implantation numbers were improved, and the degree of fibrosis was significantly alleviated by UC-MSCs treatment in the rat model of thin endometrium. In vitro cell experiments showed that UC-MSCs migrated to injured ESCs and enhanced their proliferation. miRNA microarray chip results showed that expression of 45 miRNAs was downregulated in the injured endometrium and upregulated after UC-MSCs transplantation. Likewise, expression of 39 miRNAs was upregulated in the injured endometrium and downregulated after UC-MSCs transplantation. The miRNA-mRNA interactions showed the changes in the miRNA and mRNA network during the processes of endometrial injury and repair. GO and KEGG analyses showed that the process of endometrial injury was mainly attributed to the decomposition of the extracellular matrix (ECM), protein degradation and absorption, and accompanying inflammation. The process of UC-MSCs transplantation and repair were accompanied by the reconstruction of the ECM, regulation of chemokines and inflammation, and cell proliferation and apoptosis. The key molecules involved in ECM-receptor interaction pathways were further verified by qRT-PCR. Itga1 and Thbs expression decreased in the model group and increased by UC-MSCs transplantation, while Laminin and Collagen expression increased in both the model group and MSCs group, with greater expression observed in the latter. This study showed that UC-MSCs transplantation could promote recovery of thin endometrial morphology and function. Furthermore, it revealed the expression changes of miRNA and mRNA after endometrial injury and UC-MSCs transplantation repair processed, and signaling pathways that may be involved in endometrial injury and repair.

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Influence of Chondrocyte Zone on Co-Cultures With Mesenchymal Stem Cells in HA Hydrogels for Cartilage Tissue Engineering

ASME 2012 Summer Bioengineering Conference, Parts A and B ◽

10.1115/sbc2012-80859 ◽

2012 ◽

Cited By ~ 2

Author(s):

Minwook Kim ◽

Jason A. Burdick ◽

Robert L. Mauck

Keyword(s):

Tissue Engineering ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Hyaluronic Acid ◽

Articular Chondrocytes ◽

Cartilage Tissue Engineering ◽

3D Culture ◽

Cartilage Tissue ◽

Cell Type

Mesenchymal stem cells (MSCs) are an attractive cell type for cartilage tissue engineering in that they can undergo chondrogenesis in a variety of 3D contexts [1]. Focused efforts in MSC-based cartilage tissue engineering have recently culminated in the formation of biologic materials possessing biochemical and functional mechanical properties that match that of the native tissue [2]. These approaches generally involve the continuous or intermittent application of pro-chondrogenic growth factors during in vitro culture. For example, in one recent study, we showed robust construct maturation in MSC-seeded hyaluronic acid (HA) hydrogels transiently exposed to high levels of TGF-β3 [3]. Despite the promise of this approach, MSCs are a multipotent cell type and retain a predilection towards hypertrophic phenotypic conversion (i.e., bone formation) when removed from a pro-chondrogenic environment (e.g., in vivo implantation). Indeed, even in a chondrogenic environment, many MSC-based cultures express pre-hypertrophic markers, including type X collagen, MMP13, and alkaline phosphatase [4]. To address this issue, recent studies have investigated co-culture of human articular chondrocytes and MSCs in both pellet and hydrogel environments. Chondrocytes appear to enhance the initial efficiency of MSC chondrogenic conversion, as well as limit hypertrophic changes in some instances (potentially via secretion of PTHrP and/or other factors) [5–7]. While these findings are intriguing, articular cartilage has a unique depth-dependent morphology including zonal differences in chondrocyte identity. Ng et al. showed that zonal chondrocytes seeded in a bi-layered agarose hydrogel construct can recreate depth-dependent cellular and mechanical heterogeneity, suggesting that these identities are retained with transfer to 3D culture systems [8]. Further, Cheng et al. showed that differences in matrix accumulation and hypertrophy in zonal chondrocytes was controlled by bone morphogenic protein [9]. To determine whether differences in zonal chondrocyte identity influences MSC fate decisions, we evaluated functional properties and phenotypic stability in photocrosslinked hyaluronic acid (HA) hydrogels using distinct, zonal chondrocyte cell fractions co-cultured with bone marrow derived MSCs.

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The influence of degradation characteristics of hyaluronic acid hydrogels on in vitro neocartilage formation by mesenchymal stem cells

Biomaterials ◽

10.1016/j.biomaterials.2009.04.040 ◽

2009 ◽

Vol 30 (26) ◽

pp. 4287-4296 ◽

Cited By ~ 145

Author(s):

Cindy Chung ◽

Michael Beecham ◽

Robert L. Mauck ◽

Jason A. Burdick

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Hyaluronic Acid

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Applying vibration in early postmenopausal osteoporosis promotes osteogenic differentiation of bone marrow-derived mesenchymal stem cells and suppresses postmenopausal osteoporosis progression

Bioscience Reports ◽

10.1042/bsr20191011 ◽

2019 ◽

Vol 39 (9) ◽

Cited By ~ 2

Author(s):

Huiming Li ◽

Wenchao Wu ◽

Xueling He ◽

Chengjian Cao ◽

Xiaoqin Yu ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Osteogenic Differentiation ◽

Postmenopausal Osteoporosis ◽

Biomechanical Properties ◽

Estradiol Benzoate ◽

Once Daily ◽

Underlying Mechanisms

Abstract We aimed to evaluate whether applying low magnitude vibration (LMV) in early postmenopausal osteoporosis (PMO) suppresses its progression, and to investigate underlying mechanisms. Rats were randomly divided into Sham (Sham-operated), Sham+V, OVX (ovariectomized), OVX+E2 (estradiol benzoate), OVX+V (LMV at 12–20 weeks postoperatively), and OVX+Vi (LMV at 1–20 weeks postoperatively) groups. LMV was applied for 20 min once daily for 5 days weekly. V rats were loaded with LMV at 12–20 weeks postoperatively. Vi rats were loaded with LMV at 1–20 weeks postoperatively. Estradiol (E2) rats were intramuscularly injected at 12–20 weeks postoperatively once daily for 3 days. The bone mineral densities (BMDs), biomechanical properties, and histomorphological parameters of tibiae were analyzed. In vitro, rat bone marrow-derived mesenchymal stem cells (rBMSCs) were subjected to LMV for 30 min daily for 5 days, or 17β-E2 with or without 1-day pretreatment of estrogen receptor (ER) inhibitor ICI 182,780 (ICI). The mRNA and protein expresion were performed. Data showed that LMV increased BMD, bone strength, and bone mass of rats, and the effects of Vi were stronger than those of E2. In vitro, LMV up-regulated the mRNA and protein expressions of Runx2, Osx, Col I, and OCN and down-regulated PPARγ, compared with E2. The effects of both LMV and E2 on rBMSCs were inhibited by ICI. Altogether, LMV in early PMO suppresses its progression, which is associated with osteogenic differentiation of rBMSCs via up-regulation of ERα and activation of the canonical Wnt pathway. LMV may therefore be superior to E2 for the suppression of PMO progression.

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Differential Chondrogenic Potential of Human and Bovine Mesenchymal Stem Cells in Agarose and Photocrosslinked Hyaluronic Acid Hydrogels

ASME 2010 Summer Bioengineering Conference, Parts A and B ◽

10.1115/sbc2010-19461 ◽

2010 ◽

Author(s):

Minwook Kim ◽

Isaac E. Erickson ◽

Jason A. Burdick ◽

George R. Dodge ◽

Robert L. Mauck

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Hyaluronic Acid ◽

Strong Dependence ◽

Cartilage Tissue Engineering ◽

Cartilage Tissue ◽

Human Mscs ◽

Biologically Relevant

Articular cartilage has a limited regenerative capacity, and there exist no methodologies to restore structure and function after damage or degeneration. This has focused intense work on cell-based therapies for cartilage repair, with considerable literature demonstrating that chondrocytes in vitro and in vivo can generate cartilage-like tissue replacements. However, use of primary cells is limited by the amount and quality of autologous donor cells and tissue. Multipotential mesenchymal stem cells (MSCs) derived from bone marrow offer an alternative cell source for cartilage tissue engineering. MSCs are easily accessible and expandable in culture, and differentiate towards a chondrocyte-like phenotype with exposure to TGF-β [1]. For example, we have shown that bovine MSCs undergo chondrogenic differentiation and mechanical maturation in agarose, self-assembling peptide, and photocrosslinkable hyaluronic acid (HA) hydrogels [2]. HA hydrogels are particularly advantageous as they are biologically relevant and easily modified to generate a range of hydrogel properties [3]. Indeed, bovine MSCs show a strong dependence of functional outcomes on the macromer density of the HA gel [4]. To further the clinical application of this material, the purpose of this study was to investigate functional chondrogenesis of human MSCs in HA compared to agarose hydrogels. To carry out this study, juvenile bovine and human MSCs were encapsulated and cultured in vitro in HA and agarose hydrogels, and cell viability, biochemical, biomechanical, and histological properties were evaluated over 4 weeks of culture.

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Silk fibroin/gelatin–chondroitin sulfate–hyaluronic acid effectively enhances in vitro chondrogenesis of bone marrow mesenchymal stem cells

Materials Science and Engineering C ◽

10.1016/j.msec.2015.03.043 ◽

2015 ◽

Vol 52 ◽

pp. 90-96 ◽

Cited By ~ 33

Author(s):

Nopporn Sawatjui ◽

Teerasak Damrongrungruang ◽

Wilairat Leeanansaksiri ◽

Patcharee Jearanaikoon ◽

Suradej Hongeng ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Hyaluronic Acid ◽

Chondroitin Sulfate ◽

Silk Fibroin ◽

Marrow Mesenchymal Stem Cells

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Improvement of Distribution and Osteogenic Differentiation of Human Mesenchymal Stem Cells by Hyaluronic Acid and β-Tricalcium Phosphate-Coated Polymeric Scaffold In Vitro

BioResearch Open Access ◽

10.1089/biores.2015.0021 ◽

2015 ◽

Vol 4 (1) ◽

pp. 363-373 ◽

Cited By ~ 16

Author(s):

Muwan Chen ◽

Dang Q.S. Le ◽

Jørgen Kjems ◽

Cody Bünger ◽

Helle Lysdahl

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Hyaluronic Acid ◽

Osteogenic Differentiation ◽

Tricalcium Phosphate ◽

Human Mesenchymal Stem Cells ◽

Polymeric Scaffold

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Melatonin Enhances Osteoblastogenesis from Senescent Mesenchymal Stem Cells via MMSET Mediated Chromatin Remodeling

10.21203/rs.3.rs-130548/v1 ◽

2021 ◽

Author(s):

Zhiqiang Liu ◽

Ying Xie ◽

Na Han ◽

Sheng Wang ◽

Xuelei Wei ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Osteogenic Potential ◽

Melatonin Treatment ◽

Large Numbers ◽

Elevated Expression ◽

Bone Marrow Plasma ◽

Underlying Mechanisms

Abstract Large numbers of elderly people have aging-associated osteoporosis, but efficient approaches to ameliorate bone loss are limited due to our poor understanding of the underlying mechanisms. In this study, we found that melatonin levels in bone marrow decreased with age, and melatonin primarily enhanced the osteogenic potential of mesenchymal stem cells (MSCs) derived from elderly donors compared with fetal- or young adult-derived MSCs. Mechanistic studies indicated melatonin treatment alleviated the senescence-related hypermethylation of the MMSET promoter, leading to elevated expression of the histone methyltransferase NSD2, and promoted the histone H3 dimethylation modification at lysine 36 of the osteogenic genes RUNX2 and SP7/OSTERIX as a consequence. MMSET depletion partially abolished the effects of melatonin on osteogenesis in senescent MSCs in vitro. Moreover, melatonin treatment promoted bone formation and alleviated the progression of osteoporosis in a mouse model of aging. Clinically, severity of senile osteoporosis (SOP) in patients was associated with melatonin levels in bone marrow plasma and the MMSET expression in MSCs, and melatonin treatment enhanced osteoblastogenesis from MSCs derived from SOP patients. Our study discovered a previously unreported epigenetic regulatory role for melatonin in alleviating MSC senescence and suggests that melatonin may be a potent agent for preventing aging-associated osteoporosis.

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