Tracing immune cells around biomaterials with spatial anchors during large-scale wound regeneration.

Skin scarring devoid of dermal appendages after severe trauma has unfavorable effects on aesthetic and physiological functions. Here we present a method for large-area wound regeneration using biodegradable aligned extracellular matrix scaffolds. We show that the implantation of these scaffolds accelerates wound coverage and enhances hair follicle neogenesis. We perform multimodal analysis, in combination with single-cell RNA sequencing and spatial transcriptomics, to explore the immune responses around biomaterials, highlighting the potential role of regulatory T cells in mitigating tissue fibrous by suppressing excessive type 2 inflammation. We find that immunodeficient mice lacking mature T lymphocytes show the typical characteristic of tissue fibrous driven by type 2 macrophage inflammation, validating the potential therapeutic effect of the adaptive immune system activated by biomaterials. These findings contribute to our understanding of the coordination of immune systems in wound regeneration and facilitate the design of immunoregulatory biomaterials in the future.

Macrophage response mediated by extracellular matrix: recent progress.

Biomaterials are one of efficient treatment options for tissue defects in regenerative medicine. Compared to synthetic materials which tend to induce chronic inflammatory response and fibrous capsule, extracellular matrix (ECM) scaffold materials composed of biopolymers are thought to be capable of inducing a pro-regenerative immune microenvironment and facilitate wound healing. Immune cells are the first line of response to implanted biomaterials. In particular, macrophages greatly affect cell behavior and the ultimate treatment outcome based on multiple cell phenotypes with various functions. The macrophage polarization status is considered as a general reflection of the characteristics of the immune microenvironment. Since numerous reports has emphasized the limitation of classical M1/M2 nomenclature, high-resolution techniques such as single-cell sequencing has been applied to recognize distinct macrophage phenotypes involved in host responses to biomaterials. After reviewing latest literatures that explored the immune microenvironment mediated by ECM scaffolds, this paper describe the behaviors of highly heterogeneous and plastic macrophages subpopulations which affect the tissue regeneration. The mechanisms by which ECM scaffolds interact with macrophages are also discussed from the perspectives of the ECM ultrastructure along with the nucleic acid, protein, and proteoglycan compositions, in order to provide targets for potential therapeutic modulation in regenerative medicine.

Immunomodulatory Biomaterials and Emerging Analytical Techniques for Probing the Immune Micro-Environment.

After implantation of a biomaterial, both the host immune system and properties of the material determine the local immune response. Through triggering or modulating the local immune response, materials can be designed towards a desired direction of promoting tissue repair or regeneration. High-throughput sequencing technologies such as single-cell RNA sequencing (scRNA-seq) emerging as a powerful tool for dissecting the immune micro-environment around biomaterials, have not been fully utilized in the field of soft tissue regeneration. In this review, we first discussed the procedures of foreign body reaction in brief. Then, we summarized the influences that physical and chemical modulation of biomaterials have on cell behaviors in the micro-environment. Finally, we discussed the application of scRNA-seq in probing the scaffold immune micro-environment and provided some reference to designing immunomodulatory biomaterials. The foreign body response consists of a series of biological reactions. Immunomodulatory materials regulate immune cell activation and polarization, mediate divergent local immune micro-environments and possess different tissue engineering functions. The manipulation of physical and chemical properties of scaffolds can modulate local immune responses, resulting in different outcomes of fibrosis or tissue regeneration. With the advancement of technology, emerging techniques such as scRNA-seq provide an unprecedented understanding of immune cell heterogeneity and plasticity in a scaffold-induced immune micro-environment at high resolution. The in-depth understanding of the interaction between scaffolds and the host immune system helps to provide clues for the design of biomaterials to optimize regeneration and promote a pro-regenerative local immune micro-environment.

Application of biomaterials in periodontal tissue repair and reconstruction in the presence of inflammation under periodontitis through the foreign body response: Recent progress and perspectives.

Periodontitis would cause dental tissue damage locally. Biomaterials substantially affect the surrounding immune microenvironment through treatment-oriented local inflammatory remodeling in dental periodontitis. This remodeling process is conducive to wound healing and periodontal tissue regeneration. Recent progress in understanding the foreign body response (FBR) and immune regulation, including cell heterogeneity, and cell-cell and cell-material interactions, has provided new insights into the design criteria for biomaterials applied in treatment of periodontitis. This review discusses recent progress and perspectives in the immune regulation effects of biomaterials to augment or reconstruct soft and hard tissue in an inflammatory microenvironment of periodontitis.

Application of Epigallocatechin-3-gallate (EGCG) Modified 1-Ethyl-3-(3-dimethylaminopropylcarbodiimide hydrochloride/N-hydroxy-succinimide (EDC/NHS) Cross-Linked Collagen Membrane to Promote Macrophage Adhesion.

The chemically cross-linking 1-ethyl-3-(3-dimethylaminopropylcarbodiimide hydrochloride/N-hydroxy-succinimide (EDC/NHS) collagen membrane endows such natural polymers with promising mechanical properties. Nevertheless, it is inadequate to advance the modulation of foreign body response (FBR) after implantation or guidance of tissue regeneration. In previous research, macrophages have a strong regulatory effect on regeneration, and such enhanced membranes underwent the modification with Epigallocatechin-3-gallate (EGCG) could adjust the recruitment and phenotypes of macrophages. Accordingly, we develop EGCG-EDC/NHS membranes, prepared with physical immersion, while focusing on the surface morphology through SEM, the biological activity of collagen was determined by FTIR, the activity and adhesion of cell culture in vitro, angiogenesis and monocyte/macrophage recruitment after subcutaneous implantation in vivo, are characterized. It could be concluded that it is hopeful EGCG-EDC/NHS collagen membrane can be used in implant dentistry for it not only retains the advantages of the collagen membrane itself, but also improves cell viability, adhesion, vascularization, and immunoregulation tendency.

Dissecting the microenvironment around biosynthetic scaffolds in murine skin wound healing.

The structural properties of biomaterials play crucial roles in guiding cell behavior and influencing immune responses against the material. We fabricated electrospun membranes with three types of surface topography (random, aligned, and latticed), introduced them to dorsal skin excisional wounds in mice and rats, and evaluated their effects on wound healing and immunomodulatory properties. An overview of different immune cells in the microenvironment with the help of single-cell RNA sequencing revealed diverse cellular heterogeneity in vivo. The time course of immune response was advanced toward an adaptive immunity-dominant stage by the aligned scaffold. In mice without mature T lymphocytes, lack of wound-induced hair neogenesis indicated a regulatory role of T cells on hair follicle regeneration. The microenvironment around scaffolds involved an intricate interplay of immune and cutaneous cells.

Modulation of foreign body reaction and macrophage phenotypes concerning microenvironment.

The foreign body reaction (FBR) is described as a local chronic inflammation after implantation of biomaterials in which macrophages involved intimately. At the stage of acute inflammation, mast cells release histamine, Interleukin-4 (IL-4) and Interleukin-13 (IL-13), enhancing recruitment, and fusion of macrophages in the following phase. As for chronic intensive inflammation, degradation of biomaterials would be promoted by macrophage-derived foreign body giant cells releasing degradative enzymes, acid and reactive oxygen intermediates. Nevertheless, it could be seen as a breakthrough point for regulating FBR, considering the dominant role of the macrophage in the immune response as exemplified by the decrease of IL-4 and IL-13, stabilizing an appropriate balance between two macrophage phenotypes, selectively suppressing some function of macrophages, and so on. Moreover, the relationship between macrophages polarization and the development of a fibrous capsule, which increase the possibility of implantation failure, will be illustrated later. This review aims at providing readers a comprehensive understanding of FBR and its correlative treatment strategy.

Evaluation of epigallocatechin-3-gallate (EGCG)-modified scaffold determines macrophage recruitment.

Biomaterials based on the modulation of macrophages have gained increased attention recently. Macrophages are generally divided into the pro-inflammatory M1 and pro-regenerative M2 phenotypes. Macrophages play a pivotal role in bone regeneration by regulating osteoblast differentiation and secreting pro-regenerative factors. In the present study, epigallocatechin-3-gallate (EGCG)-modified collagen membranes downregulated the expression of inflammatory factors and promoted the recruitment of M2 macrophages, as evidenced by the expression of M2 macrophage markers (CD163 and CD206). It is further demonstrated that the recruitment of M2 macrophages may be involved with CC chemokine receptor type 2 (CCR2) signaling, with a significant downregulation of CD206 following CCR2 knockout. These results suggested that EGCG-modified collagen membranes may modulate the recruitment of macrophages and can be applied to guided bone regeneration and guided tissue regeneration.

Evaluation of epigallocatechin-3-gallate (EGCG) modified collagen in guided bone regeneration (GBR) surgery and modulation of macrophage phenotype.

Collagen membranes have been widely applied for guided bone regeneration (GBR), a technique often utilized in dental implant surgery for bone argumentation. However, the implantation of collagen membranes also elicits foreign body reaction (FBR), the imbalance of which may lead to failures of dental implants. Macrophages play a pivotal role in FBR as macrophages can polarize into pro-inflammatory (M1) and pro-regenerative (M2) phenotypes. Therefore, collagen membranes based on modulation of macrophage polarization have gained increased attention. Epigallocatechin-3-gallate (EGCG)-modified collagen membranes have been previously shown to downregulate the expression of inflammatory factors. In the present study, scanning electron microscopy images showed that EGCG-modified collagen membranes prevented the migration of keratinocytes and maintained space for osteoblasts. CCK-8 and live/dead cell assays showed that EGCG-modified collagen membranes unaffected the cell viability of osteoblasts. In addition, immunofluorescent staining and quantitative real-time polymerase chain reaction showed an increased number of M2 macrophages, an upregulated expression of growth factors including vascular endothelial growth factor, bone morphogenetic protein 2, and an upregulation of osteogenic differentiation-related factors including Runt-related transcription factor 2 and osteopontin after implantation of EGCG-modified collagen membranes. Hematoxylin-eosin staining and Micro-CT further demonstrated that the application of EGCG-modified collagen membranes promoted new bone formation in vivo. From these findings it is concluded that EGCG-modified collagen membranes have promising potentials in GBR surgery which served as suitable barrier membranes and promoted bone regeneration in vivo by recruiting M2 macrophages, promoting secretion of growth factors and osteogenic differentiation.