Highly Aligned Ternary Nanofiber Matrices Loaded with MXene Expedite Regeneration of Volumetric Muscle Loss.

Current therapeutic approaches for volumetric muscle loss (VML) face challenges due to limited graft availability and insufficient bioactivities. To overcome these limitations, tissue-engineered scaffolds have emerged as a promising alternative. In this study, we developed aligned ternary nanofibrous matrices comprised of poly(lactide-co-ε-caprolactone) integrated with collagen and Ti3C2Tx MXene nanoparticles (NPs) (PCM matrices), and explored their myogenic potential for skeletal muscle tissue regeneration. The PCM matrices demonstrated favorable physicochemical properties, including structural uniformity, alignment, microporosity, and hydrophilicity. In vitro assays revealed that the PCM matrices promoted cellular behaviors and myogenic differentiation of C2C12 myoblasts. Moreover, in vivo experiments demonstrated enhanced muscle remodeling and recovery in mice treated with PCM matrices following VML injury. Mechanistic insights from next-generation sequencing revealed that MXene NPs facilitated protein and ion availability within PCM matrices, leading to elevated intracellular Ca2+ levels in myoblasts through the activation of inducible nitric oxide synthase (iNOS) and serum/glucocorticoid regulated kinase 1 (SGK1), ultimately promoting myogenic differentiation via the mTOR-AKT pathway. Additionally, upregulated iNOS and increased NO- contributed to myoblast proliferation and fiber fusion, thereby facilitating overall myoblast maturation. These findings underscore the potential of MXene NPs loaded within highly aligned matrices as therapeutic agents to promote skeletal muscle tissue recovery.

Clear Cell Adenocarcinoma of Urethra: Clinical and Pathologic Implications and Characterization of Molecular Aberrations.

PURPOSE: This study aimed to evaluate the molecular features of clear cell adenocarcinoma (CCA) of the urinary tract and investigate its pathogenic pathways and possible actionable targets. MATERIALS AND METHODS: We retrospectively collected the data of patients with CCA between January 1999 and December 2016; the data were independently reviewed by two pathologists. We selected five cases of urinary CCA, based on the clinicopathological features. We analyzed these five cases by whole exome sequencing (WES) and subsequent bioinformatics analyses to determine the mutational spectrum and possible pathogenic pathways. RESULTS: All patients were female with a median age of 62 years. All tumors were located in the urethra and showed aggressive behavior with disease progression. WES revealed several genetic alterations, including driver gene mutations (AMER1, ARID1A, CHD4, KMT2D, KRAS, PBRM1, and PIK3R1) and mutations in other important genes with tumor-suppressive and oncogenic roles (CSMD3, KEAP1, SMARCA4, and CACNA1D). We suggest putative pathogenic pathways (chromatin remodeling pathway, mitogen-activated protein kinase signaling pathway, phosphoinositide 3-kinase/AKT/mammalian target of rapamycin pathway, and Wnt/ß-catenin pathway) as candidates for targeted therapies. CONCLUSION: Our findings shed light on the molecular background of this extremely rare tumor with poor prognosis and can help improve treatment options.

3D bioprinting of human mesenchymal stem cells-laden hydrogels incorporating MXene for spontaneous osteodifferentiation.

Contemporary advances in three-dimensional (3D) bioprinting technologies have enabled the fabrication of tailored live 3D tissue mimetics. Furthermore, the development of advanced bioink materials has been highlighted to accurately reproduce the composition of a native extracellular matrix and mimic the intrinsic properties of laden cells. Recent research has shown that MXene is one of promising nanobiomaterials with osteogenic activity for bone grafts and scaffolds due to its unique atomic structure of three titanium layers between two carbon layers. In this study, the MXene-incorporated gelatin methacryloyl (GelMA) and hyaluronic acid methacryloyl (HAMA) (i.e., GelMA/HAMA-MXene) bioinks were prepared to explore if they have the potential to enable the spontaneous osteodifferentiation of human mesenchymal stem cells (hMSCs) when the hMSCs-laden GelMA/HAMA-MXene bioinks were 3D printed. The physicochemical and rheological characteristics of the GelMA/HAMA-MXene hydrogels were proven to be unprecedentedly favorable supportive matrices suited for the growth and survival of hMSCs. Furthermore, hMSCs were shown to spontaneously differentiate into osteoblasts within GelMA-HAMA/MXene composites to provide favorable microenvironments for osteogenesis. Therefore, our results suggest that the remarkable biofunctional advantages of the MXene-incorporated GelMA/HAMA bioink can be utilized in a wide range of strategies for the development of effective scaffolds in bone tissue regeneration.

Cellulose Nanofiber Films and Their Vibration Energy Harvesting.

Cellulose, the most abundant sustainable material on Earth, has excellent mechanical and physical properties, high optical transparency, biocompatibility, and piezoelectricity. So, it has many possibilities for future materials, and many researchers are interested in its application. In this paper, cellulose nanofiber (CNF) and CNF/polyvinyl alcohol (PVA) films are made, and their vibration energy harvesting is studied. CNF was isolated by chemical and physical methods, and the CNF suspension was cast on a flat substrate to make a film. A cast CNF wet film stayed in a 5 Tesla superconductor magnet for 7 days, which resulted in CNF alignment perpendicular to the magnetic field. To further improve the mechanical properties of the CNF film, mechanical stretching was applied. The CNF suspension was mixed with PVA, giving the film toughness. The cast CNF/PVA wet film was mechanically stretched and dried, which improved the CNF alignment. The fabricated CNF and CNF/PVA films were characterized using scanning electron microscopy and X-ray diffraction to verify the alignment. By stretching, the aligned CNF/PVA film exhibits the largest mechanical properties along the aligned direction. The maximum Young's modulus and tensile strength of the 50% stretched CNF/PVA film are 14.9 GPa and 170.6 MPa, respectively. Finally, a vibration energy harvesting experiment was performed by invoking the piezoelectric behavior of the pure CNF, and 50% stretched CNF/PVA films. The harvester structure was innovated by adopting a cymbal structure, which was beneficial to producing large in-plane strain on the films. The designed cymbal structure was analyzed using ANSYS, and its natural frequency was experimentally verified. The CNF/PVA film performs better vibration energy harvesting than the pure CNF film. The CNF/PVA film is applicable for biocompatible and flexible vibration energy harvesting.

Ternary MXene-loaded PLCL/collagen nanofibrous scaffolds that promote spontaneous osteogenic differentiation.

Conventional bioinert bone grafts often have led to failure in osseointegration due to low bioactivity, thus much effort has been made up to date to find alternatives. Recently, MXene nanoparticles (NPs) have shown prominent results as a rising material by possessing an osteogenic potential to facilitate the bioactivity of bone grafts or scaffolds, which can be attributed to the unique repeating atomic structure of two carbon layers existing between three titanium layers. In this study, we produced MXene NPs-integrated the ternary nanofibrous matrices of poly(L-lactide-co-ε-caprolactone, PLCL) and collagen (Col) decorated with MXene NPs (i.e., PLCL/Col/MXene), as novel scaffolds for bone tissue engineering, via electrospinning to explore the potential benefits for the spontaneous osteogenic differentiation of MC3T3-E1 preosteoblasts. The cultured cells on the physicochemical properties of the nanofibrous PLCL/Col/MXene-based materials revealed favorable interactions with the supportive matrices, highly suitable for the growth and survival of preosteoblasts. Furthermore, the combinatorial ternary material system of the PLCL/Col/MXene nanofibers obviously promoted spontaneous osteodifferentiation with positive cellular responses by providing effective microenvironments for osteogenesis. Therefore, our results suggest that the unprecedented biofunctional advantages of the MXene-integrated PLCL/Col nanofibrous matrices can be expanded to a wide range of strategies for the development of effective scaffolds in bone tissue regeneration.

3D Printing of Skin Equivalents with Hair Follicle Structures and Epidermal-Papillary-Dermal Layers Using Gelatin/Hyaluronic Acid Hydrogels.

Recent advances in three-dimensional (3D) bioprinting technologies have enabled the fabrication of sophisticated live 3D tissue analogs. Despite the existing hydrogel-based bioinks, the development of advanced bioink materials that can accurately reproduce the composition of a native extracellular matrix and mimic the intrinsic properties of laden cells remains challenging. In this study, 3D printed skin equivalents incorporating hair follicle structures and epidermal/papillary dermal layers were fabricated using gelatin methacryloyl/hyaluronic acid methacryloyl (GelMA/HAMA) bioink. The composition of collagen and glycosaminoglycan in native skin was recapitulated by adjusting the combination of GelMA and HAMA. The GelMA/HAMA bioink exhibited excellent viscoelastic and physicochemical properties, 3D printability, cytocompatibility, and functionality to maintain hair-inductive potency while facilitating spontaneous hair pore development. The results indicate that GelMA/HAMA hydrogels are promising candidates as bioinks for the 3D printing of skin equivalents. Furthermore, they may serve as useful models for skin tissue engineering and regeneration.

Population genetic analysis reveals secondary contact between Eriocheir sinensis and E. japonica in South Korea.

BACKGROUND: The Chinese mitten crab (Eriocheir sinensis) and the Japanese mitten crab (E. japonica) of the family Varunidae, which are also critical fishery species in their native habitats, including China, Japan, and South Korea, exhibit a rare migration life history for the decapod life cycle. Eriocheir sinensis and E. japonica in South Korea may have originated from speciation or secondary contact in South Korea after speciation of these two species; however, the genetic relationship between these South Korean populations remains unclear, and need to be clarified. OBJECTIVE: This study examined the population genetic properties of E. sinensis and E. japonica in South Korea to reveal the origin of their co-existence. METHODS: Mitochondrial DNA cytochrome c oxidase I sequences of 120 E. sinensis and E. japonica from 6 sampling sites in South Korea were sequenced. Their genetic diversity and haplotype data were compared to previously reported data from the Chinese mainland, Taiwan, Japan, and Russia. RESULTS: The haplotype network, analysis of molecular variance, FST values, Mantel test, and Bayesian skyline plot results of South Korean E. sinensis and E. japonica demonstrated that E. sinensis and E. japonica were isolated from their central populations, Chinese and Japanese populations, by the distance due to the temperature, salinity, ocean current flow, or a geohistorical event. In addition, genetic analysis indicated that the South Korean populations of the two species were marginal populations, exhibiting low genetic diversity, significant negative neutrality values, and radial haplotype networks. The current results also suggest that the Geum River is an overlapping habitat for the natural populations of E. sinensis and E. japonica in South Korea. Further research is required to examine the geohistorical and evolutionary event between E. sinensis and E. japonica in South Korea. CONCLUSIONS: The co-existence of E. sinensis and E. japonica in South Korea may have originated from secondary contact after their speciation.

Differential Toxicity of Graphene Family Nanomaterials Concerning Morphology.

Graphene family nanomaterials (GFNs) are well-known carbonaceous materials, which find application in several fields like optoelectronics, photocatalysis, nanomedicine, and tissue regeneration. Despite possessing many advantages in biomedical applications, GFNs exhibited toxicity depending on various parameters including dosage, size, exposure time, and kinds of administration. GFNS are majorly classified into nanosheets, quantum dots, nanoplatelets, and nanoribbons based on morphology. Understanding the toxic effects of GFNs would provide new suggestions as to how the materials can be utilized effectively. Hence, we are summarizing here some of the recent findings in cellular and animal level toxicity studies of GFNs on the perspective of their different morphologies. Notwithstanding, we highlight progress, challenges, and new toxicological approaches to ensure biosafety of GFNs for future directions.

Functional Graphene Nanomaterials-Based Hybrid Scaffolds for Osteogenesis and Chondrogenesis.

With the emerging trends and recent advances in nanotechnology, it has become increasingly possible to overcome current hurdles for bone and cartilage regeneration. Among the wide type of nanomaterials, graphene (G) and its derivatives (graphene-based materials, GBMs) have been highlighted due to the specific physicochemical and biological properties. In this review, we present the recent development of GBM-based scaffolds for bone and cartilage engineering, focusing on the formulation/shape/size-dependent characteristics, types of scaffold and modification, biocompatibility, bioactivity and underlying mechanism, drawback and prospect of each study. From the findings described herein, mechanical property, biocompatibility, osteogenic and chondrogenic property of GBM-based scaffolds could be significantly enhanced through various scaffold fabrication methods and conjugation with polymers/nanomaterials/drugs. In conclusion, the results presented in this review support the promising prospect of using GBM-based scaffolds for improved bone and cartilage tissue engineering. Although GBM-based scaffolds have some limitations to be overcome by future research, we expect further developments to provide innovative results and improve their clinical potential for bone and cartilage regeneration.

Potential of Carbon-Based Nanocomposites for Dental Tissue Engineering and Regeneration.

While conventional dental implants focus on mechanical properties, recent advances in functional carbon nanomaterials (CNMs) accelerated the facilitation of functionalities including osteoinduction, osteoconduction, and osseointegration. The surface functionalization with CNMs in dental implants has emerged as a novel strategy for reinforcement and as a bioactive cue due to their potential for mechanical reinforcing, osseointegration, and antimicrobial properties. Numerous developments in the fabrication and biological studies of CNMs have provided various opportunities to expand their application to dental regeneration and restoration. In this review, we discuss the advances in novel dental implants with CNMs in terms of tissue engineering, including material combination, coating strategies, and biofunctionalities. We present a brief overview of recent findings and progression in the research to show the promising aspect of CNMs for dental implant application. In conclusion, it is shown that further development of surface functionalization with CNMs may provide innovative results with clinical potential for improved osseointegration after implantation.

Recent Trends in Exhaled Breath Diagnosis Using an Artificial Olfactory System.

Artificial olfactory systems are needed in various fields that require real-time monitoring, such as healthcare. This review introduces cases of detection of specific volatile organic compounds (VOCs) in a patient's exhaled breath and discusses trends in disease diagnosis technology development using artificial olfactory technology that analyzes exhaled human breath. We briefly introduce algorithms that classify patterns of odors (VOC profiles) and describe artificial olfactory systems based on nanosensors. On the basis of recently published research results, we describe the development trend of artificial olfactory systems based on the pattern-recognition gas sensor array technology and the prospects of application of this technology to disease diagnostic devices. Medical technologies that enable early monitoring of health conditions and early diagnosis of diseases are crucial in modern healthcare. By regularly monitoring health status, diseases can be prevented or treated at an early stage, thus increasing the human survival rate and reducing the overall treatment costs. This review introduces several promising technical fields with the aim of developing technologies that can monitor health conditions and diagnose diseases early by analyzing exhaled human breath in real time.

Reduced graphene oxide coating enhances osteogenic differentiation of human mesenchymal stem cells on Ti surfaces.

BACKGROUND: Titanium (Ti) has been utilized as hard tissue replacement owing to its superior mechanical and bioinert property, however, lack in tissue compatibility and biofunctionality has limited its clinical use. Reduced graphene oxide (rGO) is one of the graphene derivatives that possess extraordinary biofunctionality and are known to induce osseointegration in vitro and in vivo. In this study, rGO was uniformly coated by meniscus-dragging deposition (MDD) technique to fabricate rGO-Ti substrate for orthopedic and dental implant application. METHODS: The physicochemical characteristics of rGO-coated Ti (rGO-Ti) substrates were evaluated by atomic force microscopy, water contact angle, and Raman spectroscopy. Furthermore, human mesenchymal stem cells (hMSCs) were cultured on the rGO-Ti substrate, and then their cellular behaviors such as growth and osteogenic differentiation were determined by a cell counting kit-8 assay, alkaline phosphatase (ALP) activity assay, and alizarin red S staining. RESULTS: rGO was coated uniformly on Ti substrates by MDD process, which allowed a decrease in the surface roughness and contact angle of Ti substrates. While rGO-Ti substrates significantly increased cell proliferation after 7 days of incubation, they significantly promoted ALP activity and matrix mineralization, which are early and late differentiation markers, respectively. CONCLUSION: It is suggested that rGO-Ti substrates can be effectively utilized as dental and orthopedic bone substitutes since these graphene derivatives have potent effects on stimulating the osteogenic differentiation of hMSCs and showed superior bioactivity and osteogenic potential.

Advanced Techniques for Skeletal Muscle Tissue Engineering and Regeneration.

Tissue engineering has recently emerged as a novel strategy for the regeneration of damaged skeletal muscle tissues due to its ability to regenerate tissue. However, tissue engineering is challenging due to the need for state-of-the-art interdisciplinary studies involving material science, biochemistry, and mechanical engineering. For this reason, electrospinning and three-dimensional (3D) printing methods have been widely studied because they can insert embedded muscle cells into an extracellular-matrix-mimicking microenvironment, which helps the growth of seeded or laden cells and cell signals by modulating cell-cell interaction and cell-matrix interaction. In this mini review, the recent research trends in scaffold fabrication for skeletal muscle tissue regeneration using advanced techniques, such as electrospinning and 3D bioprinting, are summarized. In conclusion, the further development of skeletal muscle tissue engineering techniques may provide innovative results with clinical potential for skeletal muscle regeneration.

PERSISTENT REMNANTS OF DEXAMETHASONE INTRAVITREAL IMPLANT (OZURDEX).

PURPOSE: To describe persistent remnants of dexamethasone intravitreal implant (DEX implant) and evaluate the risk factors for remnant persistence in patients who received a DEX implant. METHODS: This retrospective observational study involved a retrospective chart review of 133 eyes from 129 patients who received DEX implant injection. RESULTS: Seventeen eyes of 17 patients (12.78%) exhibited remnant persistence >1 year after DEX implant injection; eight eyes (13.33%) were diagnosed with branching retinal vein occlusion, three eyes (14.28%) were diagnosed with central retinal vein occlusion, and six eyes (35.3%) were diagnosed with diabetic macular edema. The number of intravitreal DEX implant injections and triamcinolone acetonide injections administered to eyes with remnant persistence was significantly higher than that administered to the eyes without remnant persistence (P < 0.001 and P = 0.024, respectively). Remnants caused symptomatic floaters in 11.7% (2/17 eyes) of the patients with remnants and in 1.5% (2/133) of the entire patient cohort. CONCLUSION: DEX implant was not completely dissolved in the eyes that received multiple DEX implant or triamcinolone acetonide injections.

Intravitreal aflibercept for submacular hemorrhage secondary to neovascular age-related macular degeneration and polypoidal choroidal vasculopathy.

PURPOSE: To evaluate the efficacy of intravitreal aflibercept monotherapy for submacular hemorrhage secondary to neovascular age-related macular degeneration (AMD) and polypoidal choroidal vasculopathy (PCV). METHODS: This prospective, phase 4 clinical trial included 29 patients diagnosed with fovea-involving submacular hemorrhage secondary to neovascular AMD (7 patients) or PCV (22 patients). Patients were initially administered 3 monthly aflibercept injections, followed by 1 injection every 2 months. The primary outcome measure was changes in Early Treatment Diabetic Retinopathy Study (ETDRS) best-corrected visual acuity (BCVA) during the 56-week study period. Other key outcome measures were the proportion of patients who exhibited changes in BCVA of ≥ 15 ETDRS letters from baseline and changes in central retinal thickness (CRT). RESULTS: The mean size of hemorrhage was 6.2 ± 4.8-disc-diameter area. The mean BCVA significantly improved from 52.9 ± 17.8 ETDRS letters at week 0 (baseline) to 71.8 ± 16.1 letters at week 56 (P < 0.001). At week 56, improvement in BCVA of ≥ 15 letters was noted in 16 patients (55.2%), whereas none of the patients experienced a loss of ≥ 15 letters. The mean CRT significantly decreased from 498.9 ± 194.2 µm at week 0 to 248.3 ± 45.0 µm at week 56 (P < 0.001). During the study period, retinal break developed in one patient. CONCLUSIONS: Intravitreal aflibercept administered every 2 months after the 3 initial monthly doses was found to be an effective and safe treatment method for submacular hemorrhage secondary to neovascular AMD.

The complete mitochondrial genome sequence of the intertidal crab Parasesarma Tripectinis (Arthropoda, Decapoda, Sesarmidae).

Parasesarma tripectinis is known as an intertidal crab and inhabits Asian region. This crab has larval release at semilunar rhythm. Here, we report the complete sequence of the mitochondrial genome (mitogenome), which is composed of 15,612 base pair (bp) encoding 13 protein-coding genes, 22 transfer RNAs, 2 ribosomal RNAs, and an A + T rich region. The nucleotide composition of P. tripectinis was G + C: 25.8%, A + T: 74.2%, with a strong AT bias. Phylogenetic analysis using whole mitogenome figured out that P. tripectinis was closely related to Sesarma neglectum which belongs to the same family Sesarmidae.

Spatholobus suberectus Exhibits Antidiabetic Activity In Vitro and In Vivo through Activation of AKT-AMPK Pathway.

Glucose deposition in peripheral tissue is an important parameter for the treatment of type 2 diabetes mellitus. The aim of this study was to investigate the effects of Spatholobus suberectus (Ss) on glucose disposal in skeletal muscle cells and additionally explore its in vivo antidiabetic potential. Treatment of ethanolic extract of S. suberectus (EeSs) significantly enhanced the glucose uptake, mediated through the enhanced expression of GLUT4 in skeletal muscle via the stimulation of AKT and AMPK pathways in C2C12 cells. Moreover, EeSs have potential inhibitory action on α-glucosidase activity and significantly lowered the postprandial blood glucose levels in STZ-induced diabetic mice, associated with increased expression of GLUT4 and AKT and/or AMPK-mediated signaling cascade in skeletal muscle. Furthermore, administration of EeSs significantly boosted up the antioxidant enzyme expression and also mitigated the gluconeogenesis enzyme such as PEPCK and G-6-Pase enzyme expression in liver tissue of STZ-induced diabetic mice model. Collectively, these findings suggest that EeSs have a high potentiality to mitigate diabetic symptoms through stimulating glucose uptake in peripheral tissue via the activation of AKT and AMPK signaling cascade and augmenting antioxidant potentiality as well as blocking the gluconeogenesis process in diabetic mice.

Lannea coromandelica (Houtt.) Merr. Induces Heme Oxygenase 1 (HO-1) Expression and Reduces Oxidative Stress via the p38/c-Jun N-Terminal Kinase-Nuclear Factor Erythroid 2-Related Factor 2 (p38/JNK-NRF2)-Mediated Antioxidant Pathway.

The leaves of Lannea coromandelica (Houtt.) Merr. are used in the Garo, Pahan, and Teli tribal communities of Bangladesh as a traditional medicinal plant to treat hepatitis, diabetes, ulcers, heart disease, and dysentery. However, there have been limited phytochemical and biological studies on the bark of L. coromandelica. This study aimed to investigate the antioxidant activities of L. coromandelica bark extract (LCBE) and the underlying mechanism using RAW 264.7 cells. The LCBE was analysed by high-pressure liquid chromatography (HPLC) to detect its key polyphenolic compounds. Various in vitro antioxidant assays were performed using RAW 264.7 cells to assess the antioxidant effects of the LCBE and to understand the underlying molecular mechanism. HPLC revealed the presence of gallic acid, (-)-epigallocatechin-3-gallate, catechin, chlorogenic acid, and caffeic acid in the LCBE. The extract showed a very potent capacity to scavenge numerous free radicals through hydrogen atom transfer and/or electron donation and also quenched cellular reactive oxygen species (ROS) generation without showing any toxicity. The LCBE was found to combat the oxidative stress by enhancing the expression, at both transcriptional and translational levels, of primary antioxidant enzymes as well as phase II detoxifying enzymes, especially heme oxygenase 1, through the upregulation of the nuclear factor erythroid 2-related factor 2 (NRF2)-mediated pathway in RAW 264.7 cells via the phosphorylation of p38 kinase and c-Jun N-terminal kinase (JNK). The LCBE exhibited strong antioxidant activities and mitigated the cellular ROS production. These results provide scientific evidence of its potential as an ideal applicant for a cost-effective, readily available, and natural phytochemical, as well as a strategy for preventing diseases associated with oxidative stress and attenuating disease progress.

The first complete mitochondrial genome sequence of the leucosiid crab Pyrhila pisum (Arthropoda, Decapoda, Leucosiidae).

Pyrhila pisum is known as leucosiid crab. So far, there is no study about whole mitochondrial genome in Leucosiidae family. Here, we report first the complete sequence of the mitochondrial genome from P. pisum, which is composed of 15,516 base pair encoding 13 protein-coding genes, 22 transfer RNAs, two ribosomal RNAs, and A + T-rich region. The nucleotide composition of P. pisum was G + C: 25.5%, A + T: 74.5%, with a strong AT bias. In phylogenetic analysis using whole mitogenome, it was figured out that P. pisum was closely related to Sesarma neglectum but their family is different.

Evaluation of stem cell components in retrocorneal membranes.

The purpose of this study was to elucidate the origin and cellular composition of retrocorneal membranes (RCMs) associated with chemical burns using immunohistochemical staining for primitive cell markers. Six cases of RCMs were collected during penetrating keratoplasty. We examined RCMs with hematoxylin and eosin (H&E), periodic acid-Schiff (PAS) staining and immunohistochemical analysis using monoclonal antibodies against hematopoietic stem cells (CD34, CD133, c-kit), mesenchymal stem cells (beta-1-integrin, TGF-ß, vimentin, hSTRO-1), fibroblasts (FGF-ß, α-smooth muscle actin), and corneal endothelial cells (type IV collagen, CD133, VEGF, VEGFR1). Histologic analysis of RCMs revealed an organized assembly of spindle-shaped cells, pigment-laden cells, and thin collagenous matrix structures. RCMs were positive for markers of mesenchymal stem cells including beta-1-integrin, TGF-ß, vimentin, and hSTRO-1. Fibroblast markers were also positive, including FGF-ß and α-smooth muscle actin (SMA). In contrast, immunohistochemical staining was negative for hematopoietic stem cell markers including CD34, CD133 and c-kit as well as corneal endothelial cell markers such as type IV collagen, CD133 except VEGF and VEGFR1. Pigment-laden cells did not stain with any antibodies. The results of this study suggest that RCMs consist of a thin collagen matrix and fibroblast-like cells and may be a possible neogenetic structure produced from a lineage of bone marrow-derived mesenchymal stem cells.