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Mesenchymal stem cells (MSCs) that meet the International Society for Cellular Therapy (ISCT) criteria are obtained from placental tissue by plastic adherence. Historically, no known single marker was available for isolating placental MSCs (pMSCs) from the decidua basalis. As the decidua basalis is derived from the regenerative endometrium, we hypothesised that SUSD2, an endometrial perivascular MSC marker, would purify maternal perivascular pMSC. Perivascular pMSCs were isolated from the maternal placenta using SUSD2 magnetic bead sorting and assessed for the colony-forming unit-fibroblasts (CFU-F), surface markers, and in vitro differentiation into mesodermal lineages. Multi-colour immunofluorescence was used to colocalise SUSD2 and α-SMA, a perivascular marker in the decidua basalis. Placental stromal cell suspensions comprised 5.1%SUSD2+ cells. SUSD2 magnetic bead sorting of the placental stromal cells increased their purity approximately two-fold. SUSD2+ pMSCs displayed greater CFU-F activity than SUSD2- stromal fibroblasts (pSFs). However, both SUSD2+ pMSC and SUSD2- pSF underwent mesodermal differentiation in vitro, and both expressed the ISCT surface markers. Higher percentages of cultured SUSD2+ pMSCs expressed the perivascular markers CD146, CD140b, and SUSD2 than SUSD2- pSFs. These findings suggest that SUSD2 is a single marker that enriches maternal pMSCs, suggesting they may originate from eMSC. Placental decidua basalis can be used as an alternative source of MSC for clinical translation in situations where there is no access to endometrial tissue.

Mesenchymal stem cells (MSCs) that meet the International Society for Cellular Therapy (ISCT) criteria are obtained from placental tissue by plastic adherence. Historically, no known single marker was available for isolating placental MSCs (pMSCs) from the decidua basalis. As the decidua basalis is derived from the regenerative endometrium, we hypothesised that SUSD2, an endometrial perivascular MSC marker, would purify maternal perivascular pMSC. Perivascular pMSCs were isolated from the maternal placenta using SUSD2 magnetic bead sorting and assessed for the colony-forming unit-fibroblasts (CFU-F), surface markers, and in vitro differentiation into mesodermal lineages. Multi-colour immunofluorescence was used to colocalise SUSD2 and α-SMA, a perivascular marker in the decidua basalis. Placental stromal cell suspensions comprised 5.1%SUSD2

In the present study, the bone morphogenetic protein-2 (BMP-2) was loaded onto a polydopamine-coated biphasic calcium phosphate (BCP) granule system which could be effectively used as an injectable bone substitute (IBS) for improved bone regeneration. The surface of BCP granules was first coated with polydopamine (PD-MCG) and then BMP-2 growth factor was immobilized onto the PD-MCG by a conventional method. The results showed that BMP-2 was successfully immobilized into polydopamine-coated granules and about 87% of BMP-2 was retained on the granular surface for 30 days. Invitro and invivo studies showed enhanced cell viability and superior bone formation conferred by B-PD-MCG granules compared with PD-MCG and MCG. The percent healing capacity (BV/SV) of the three samples tested was found to be 1.64 ± 0.04, 1.4 ± 0.02 and 1.12 ± 0.03 for B-PD-MCG, PD-MCG and MCG, respectively.

The objective of the present study was to incorporate surface modified porous multichannel BCP granule into CPC to enhance its in vivo biodegradation and bone tissue growth. The multichannel BCP granule (15wt%) was first coated with collagen subsequent to BMP-2 loading (ccMCG-B). It was then embedded into CPC to form CPC-ccMCG-B system. The newly developed CPC-ccMCG-B system was then examined for SEM, EDX, XRD, setting time, compressive strength, injectability, pH change, BMP-2 release, in vitro as well as in vivo studies and further compared with CPC. Optimized CPC (0.45mL/g) was found based on setting time and compressive strength studies. In vivo studies exhibited improved new bone formation and better degradation of CPC after 2 and 4weeks of implantation as compared to CPC as resulted from effective BMP-2 signaling. Our results suggest that CPC-ccMCG-B system might be used as a promising injectable bone substitutes in clinical applications.

Endometrial mesenchymal stem/stromal cells (eMSCs) exhibit excellent regenerative capacity in the endometrial lining of the uterus following menstruation and high proliferative capacity in vitro. Bioprinting eMSCs onto a mesh could be a potential therapy for Pelvic Organ Prolapse (POP). This study reports a novel strategy targeting vaginal repair using bioprinting of eMSCs encapsulated in a hydrogel and3D melt electrospun mesh to generate a tissue engineering construct. Following a CAD, 3D printed poly e-caprolactone (PCL) meshes were fabricated using melt electrospinning (MES) at different temperatures using a GMP clinical grade GESIM Bioscaffolder. Electron and atomic force microscopies revealed that MES meshes fabricated at 100°C and with a speed 20 mm/sec had the largest open pore diameter (47.2 ± 11.4 μm) and the lowest strand thickness (121.4 ± 46 μm) that promoted optimal eMSC attachment. An Aloe Vera-Sodium Alginate (AV-ALG) composite based hydrogel was optimised to a 1:1 mixture (1%AV-1%ALG) and eMSCs, purified from human endometrial biopsies, were then bioprinted in this hydrogel onto the MES printed meshes. Acute in vivo foreign body response assessment in NSG mice revealed that eMSC printed on MES constructs promoted tissue integration, eMSC retention and an anti-inflammatory M2 macrophage phenotype characterised by F4/80+CD206+ colocalization. Our results address an unmet medical need highlighting the potential of alternative 3D bioprinted eMSC-MES meshes as a novel approach to overcome the current challenges with non-degradable knitted meshes in POP treatment.

Vaginal childbirth causes significant maternal tissue injury which can later leads to incurable disorders such as pelvic organ prolapse (POP). This study introduces an immediate vaginal tissue treatment using Aloe Vera (AV)-Alginate (ALG) hydrogel (Hyd) for delivering maternal endometrium derived Mesenchymal stem cells (eMSCs) to promote early healing in a rat simulated birth injury (SBI) model. Vaginal trauma after SBI was evidenced by increased vaginal diameter and inflammatory response. A lack of therapy showed significant reduction of smooth muscle content, increased elastin and increased tissue stiffness, indicative of fibrotic healing. Local injection with hydrogel (Hyd T) or with eMSC in hydrogel (Hyd/eMSC T) had significant impact on birth injury reversal. Hyd/eMSC T significantly improvement smooth muscle and elastin content, comparable to uninjured control vagina. At the nanoscopic level, injury caused disorganisation of structural collagen with increased D period but intervention with Hyd/eMSC T, normalised collagen structure and significantly reduced tissue stiffness. Hyd/eMSC injection showed significant immunomodulation evidenced by lowered M1:M2 ratio while restoring connective tissue composition after 6-weeks. Immediate treatment of severe vaginal birth trauma with therapeutic eMSCs delivered in AV-ALG hydrogel may be a potential new treatment strategy for healing birth injury and preventing future POP in women.

The present study addresses the effect of incorporation of alginate (ALG)-hyaluronic acid (HA) microbeads into calcium phosphate cement (CPC) based injectable bone substitute system. The morphology, chemical structure, setting time, compressive strength, injectability, washout resistance and in-vivo biological performance of CPC and CPC/ALG-HA were examined. Addition of ALG-HA microbeads in CPC resulted in an increase in the injectability and washout resistance of CPC. In-vivo studies of CPC/ALG-HA showed an improved bone formation after 8 weeks compared to CPC. The percent bone healing efficiency of CPC/ALG-HA and CPC was 26.6 ± 0.6 and 21.4 ± 0.5, respectively. The results suggest that CPC/ALG-HA could be used as a potential injectable bone substitute for improved bone regeneration.

Pelvic organ prolapse (POP) is a hidden women’s health disorder that impacts 1 in 4 women across all age groups. Surgical intervention has been the only treatment option, often involving non-degradable meshes, with variable results. However, recent reports have highlighted the adverse effects of meshes in the long term, which involve unacceptable rates of erosion, chronic infection and severe pain related to mesh shrinkage. Therefore, there is an urgent unmet need to fabricate of new class of biocompatible meshes for the treatment of POP. This review focuses on the causes for the downfall of commercial meshes, and discusses the use of emerging technologies such as electrospinning and 3D printing to design new meshes. Furthermore, we discuss the impact and advantage of nano-/microstructured alternative meshes over commercial meshes with respect to their tissue integration performance. Considering the key challenges of current meshes, we discuss the potential of cell-based tissue engineering strategies to augment the new class of meshes to improve biocompatibility and immunomodulation. Finally, this review highlights the future direction in designing the new class of mesh to overcome the hurdles of foreign body rejection faced by the traditional meshes, in order to have safe and effective treatment for women in the long term.

Zinc (Zn) enhances bone formation with mineralization and is an essential element of osteoblastic proliferation. Silicon (Si) is important in apatite formation coupled with the promotion of osteogenesis. The primary focus of this work was the assessment of the bone healing capacity of calcium phosphate cements (CPC) composed of Zn- and Si-incorporated β-tri calcium phosphate (TCP) and mono calcium phosphate mono hydrate (MCPM). Zn- and Si-incorporated β-TCP was synthesized through a sol gel process with varying amounts of Zn: (3, 6, or 9% w/w) and 15% w/w Si. Fabricated CPC samples were characterized by scanning electron microscopy, setting time, injectability, compressive strength and initial pH change with time. Compositional analysis and the effects of Zn and Si on cellular interaction were evaluated by energy dispersive X-ray spectroscopy mapping, viability determination and F-actin assay. The data were used to optimize the CPC formulation. The efficacy of bone healing was investigated via implantation into critical sized rabbit femoral condyle defects for 4 and 8 weeks. CPC cement with 6% (w/w) Zn content was the best candidate for faster bone healing (bone to tibial volume ratio in 8 weeks: 22.78% ± 0.02). Significantly faster degradation was also revealed. Bone healing was significantly delayed when CPC cement with 9% (w/w) Zn was used. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 260-271, 2017.

In this work, we report brushite-based calcium phosphate cement (CPC) system to enhance the in vivo biodegradation and tissue in-growth by incorporation of micro-channeled hydroxyapatite (HAp) granule and silicon and sodium addition in calcium phosphate precursor powder. Sodium- and silicon-rich calcium phosphate powder with predominantly tri calcium phosphate (TCP) phase was synthesized by an inexpensive wet chemical route to react with mono calcium phosphate monohydrate (MCPM) for making the CPC. TCP nanopowder also served as a packing filler and moderator of the reaction kinetics of the setting mechanism. Strong sintered cylindrical HAp granules were prepared by fibrous monolithic (FM) process, which is 800 µm in diameter and have seven micro-channels. Acid sodium pyrophosphate and sodium citrate solution was used as the liquid component which acted as a homogenizer and setting time retarder. The granules accelerated the degradation of the brushite cement matrix as well as improved the bone tissue in-growth by permitting an easy access to the interior of the CPC through the micro-channels. The addition of micro-channeled granule in the CPC introduced porosity without sacrificing much of its compressive strength. In vivo investigation by creating a critical size defect in the femur head of a rabbit model for 1 and 2 months showed excellent bone in-growth through the micro-channels. The granules enhanced the implant degradation behavior and bone regeneration in the implanted area was significantly improved after two months of implantation.

Blood serum contains various kinds of proteins which are necessary for tissue repair and regeneration process. Defect healing of fractured bone is initiated by the influx of blood and then clot formation. Thus, proteins in serum may have the ability to stimulate the bone regeneration process. In this work, we investigated the fabrication of serum-loaded oxidized alginate–gelatin–biphasic calcium phosphate hydrogels with various contents of blood serum (0%, 5%, 10%, and 15% in % v/v) to evaluate the stimulatory effect of serum proteins on bone regeneration. This system was also evaluated for rat bone marrow–derived stem cell delivery to get faster bone healing. The serum-loaded oxidized alginate–gelatin–biphasic calcium phosphate hydrogel samples were characterized by scanning electron microscopy, porosity meter, X-ray diffraction, and Fourier transform infrared for morphology and phase characterization together with their mechanical behavior. Protein release behavior, degradation, and swelling of the samples were studied. In vitro study was performed using bone marrow–derived stem cells to study cell attachment, viability, and proliferation. These studies revealed the best cell attachment and highest proliferation for 5% serum-loaded oxidized alginate–gelatin–biphasic calcium phosphate hydrogel scaffold. This composition also showed the ability to deliver stem cell in the defect zone which significantly improved the bone regeneration extent found in the in vivo animal model. In vivo study revealed that for the critical 5-mm calvarial defect into nude mouse skull, the 5% serum-loaded sample with bone marrow–derived stem cells shows the best bone regeneration potential.

The current study is a comparative assessment of the bone regeneration potentiality of bone substitutes composed of bioactive polymer-coated biphasic calcium phosphate (BCP) granules. The bone substitutes were primarily composed of multichanneled BCP granules, which were coated separately with biochemical polymer coatings, namely collagen, heparin, or polydopamine (PD), using chemical methods. The morphologic features and chemical structure of the granules and surface coatings were characterized by scanning electron microscopy and Fourier transform infrared spectroscopy, respectively. The biological characterizations in terms of cellular interaction with the bone substitutes in vitro (MTT assay and proliferation) and in vivo (bone regeneration efficacy) were performed and compared with uncoated BCP granules. It was found that PD-coated BCP granules were superior to the others with respect to promoting more rapid healing. Therefore, PD multichannel BCP granule system can be used as a promising injectable bone substitute for clinical applications.

An excessive foreign body response (FBR) has contributed to the adverse events associated with polypropylene mesh usage for augmenting pelvic organ prolapse surgery. Consequently, current biomaterial research considers the critical role of the FBR and now focuses on developing better biocompatible biomaterials rather than using inert implants to improve the clinical outcomes of their use. Tissue engineering approaches using mesenchymal stem cells (MSCs) have improved outcomes over traditional implants in other biological systems through their interaction with macrophages, the main cellular player in the FBR. The unique angiogenic, immunomodulatory and regenerative properties of MSCs have a direct impact on the FBR following biomaterial implantation. In this review, we focus on key aspects of the FBR to tissue-engineered MSC-based implants for supporting pelvic organs and beyond. We also discuss the immunomodulatory effects of the recently discovered endometrial MSCs on the macrophage response to new biomaterials designed for use in pelvic floor reconstructive surgery. We conclude with a focus on considerations in biomaterial design that take into account the FBR and will likely influence the development of the next generation of biomaterials for gynaecological applications.

A functionally modified peptide-conjugated hydrogel system was fabricated with oxidized alginate/gelatin loaded with biphasic calcium phosphate to improve its biocompatibility and functionality. Sodium alginate was treated by controlled oxidation to transform the cis-diol group into an aldehyde group in a controlled manner, which was then conjugated to the amine terminus of glycine–arginine–glycine–aspartic. Oxidized alginate glycine–arginine–glycine–aspartic was then combined with gelatin-loaded biphasic calcium phosphate to form a hydrogel of composite oxidized alginate/gelatin/biphasic calcium phosphate that displayed enhanced human adipose stem cell adhesion, spreading and differentiation. 1H nuclear magnetic resonance and electron spectroscopy for chemical analysis confirmed that the glycine–arginine–glycine–aspartic was successfully grafted to the oxidized alginate. Co-delivery of glycine–arginine–glycine–aspartic and human adipose stem cell in a hydrogel matrix was studied with the results indicating that hydrogel incorporated modified with glycine–arginine–glycine–aspartic and seeded with human adipose stem cell enhanced osteogenesis in vitro and bone formation in vivo.

A bipotential polyelectrolyte complex with biphasic calcium phosphate (BCP) powder dispersion provides an excellent option for protein adsorption and cell attachment and can facilitate enhanced bone regeneration. Application of the bipotential polyelectrolyte complex embedded in a spongy scaffold for faster healing of large segmental bone defects (LSBD) can be a promising endeavor in tissue engineering application. In the present study, a hollow scaffold suitable for segmental long bone replacement was fabricated by the sponge replica method applying the microwave sintering process. The fabricated scaffold was coated with calcium alginate at the shell surface, and genipin-crosslinked chitosan with biphasic calcium phosphate (BCP) dispersion was loaded at the central hollow core. The chitosan core was subsequently loaded with BMP-2. The electrolytic complex was characterized using SEM, porosity measurement, FTIR spectroscopy and BMP-2 release for 30 days. In vitro studies such as MTT, live/dead, cell proliferation and cell differentiation were performed. The scaffold was implanted into a 12 mm critical size defect of a rabbit radius. The efficacy of this complex is evaluated through an in vivo study, one and two month post implantation. BV/TV ratio for BMP-2 loaded sample was (42 +/- 1.76) higher compared with hollow BCP scaffold (32 +/- 0.225).

Zinc (Zn) enhances bone formation with mineralization and is an essential element of osteoblastic proliferation. Silicon (Si) is important in apatite formation coupled with the promotion of osteogenesis. The primary focus of this work was the assessment of the bone healing capacity of calcium phosphate cements (CPC) composed of Zn- and Si-incorporated β-tri calcium phosphate (TCP) and mono calcium phosphate mono hydrate (MCPM). Zn- and Si-incorporated β-TCP was synthesized through a sol gel process with varying amounts of Zn: (3, 6, or 9% w/w) and 15% w/w Si. Fabricated CPC samples were characterized by scanning electron microscopy, setting time, injectability, compressive strength and initial pH change with time. Compositional analysis and the effects of Zn and Si on cellular interaction were evaluated by energy dispersive X-ray spectroscopy mapping, viability determination and F-actin assay. The data were used to optimize the CPC formulation. The efficacy of bone healing was investigated via implantation into critical sized rabbit femoral condyle defects for 4 and 8 weeks. CPC cement with 6% (w/w) Zn content was the best candidate for faster bone healing (bone to tibial volume ratio in 8 weeks: 22.78% ± 0.02). Significantly faster degradation was also revealed. Bone healing was significantly delayed when CPC cement with 9% (w/w) Zn was used. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 260-271, 2017.