Early Stages Of Regeneration Research Articles

MTOR activity is essential for retinal pigment epithelium regeneration in zebrafish.

The retinal pigment epithelium (RPE) plays numerous critical roles in maintaining vision and this is underscored by the prevalence of degenerative blinding diseases like age-related macular degeneration (AMD), in which visual impairment is caused by progressive loss of RPE cells. In contrast to mammals, zebrafish possess the ability to intrinsically regenerate a functional RPE layer after severe injury. The molecular underpinnings of this regenerative process remain largely unknown yet hold tremendous potential for developing treatment strategies to stimulate endogenous regeneration in the human eye. In this study, we demonstrate that the mTOR pathway is activated in RPE cells post-genetic ablation. Pharmacological and genetic inhibition of mTOR activity impaired RPE regeneration, while mTOR activation enhanced RPE recovery post-injury, demonstrating that mTOR activity is essential for RPE regeneration in zebrafish. RNA-seq of RPE isolated from mTOR-inhibited larvae identified a number of genes and pathways dependent on mTOR activity at early and late stages of regeneration; amongst these were components of the immune system, which is emerging as a key regulator of regenerative responses across various tissue and model systems. Our results identify crosstalk between macrophages/microglia and the RPE, wherein mTOR activity is required for recruitment of macrophages/microglia to the RPE injury site. Macrophages/microglia then reinforce mTOR activity in regenerating RPE cells. Interestingly, the function of macrophages/microglia in maintaining mTOR activity in the RPE appeared to be inflammation-independent. Taken together, these data identify mTOR activity as a key regulator of RPE regeneration and link the mTOR pathway to immune responses in facilitating RPE regeneration.

Controlling the Spatiotemporal Release of Nerve Growth Factor by Chitosan/Polycaprolactone Conduits for Use in Peripheral Nerve Regeneration

Tubular polymeric structures have been recognized in the treatment of peripheral nerves as comparable to autologous grafting. The best therapeutic outcomes are obtained with conduits releasing therapeutic molecules. In this study, a new approach for the incorporation of biologically active agent-loaded microspheres into the structure of chitosan/polycaprolactone conduits was developed. The support of a polycaprolactone helix formed by 3D melt extrusion was coated with dopamine in order to adsorb nerve growth factor-loaded microspheres. The complex analysis of the influence of process factors on the coverage efficiency of polycaprolactone helix by nerve grow factor-loaded microspheres was analyzed. Thus, the PCL helix characterized with the highest adsorption of microspheres was subjected to nerve growth factor release studies, and finally incorporated into chitosan hydrogel deposit through the process of electrophoretic deposition. It was demonstrated by chemical and physical tests that the chitosan/polycaprolactone conduit meets the requirements imposed on peripheral nerve implants, particularly mimicking mechanical properties of surrounding soft tissue. Moreover, the conduit may support regrowing nerves for a prolonged period, as its structure and integrity persist upon incubation in lysozyme-contained PBS solution up to 28 days at body temperature. In vitro cytocompatibility toward mHippoE-18 embryonic hippocampal cells of the chitosan/polycaprolactone conduit was proven. Most importantly, the developed conduits stimulate axonal growth and support monocyte activation, the latter is advantageous especially at early stages of nerve regeneration. It was demonstrated that, through the described approach for controlling spatiotemporal release of nerve growth factors, these biocompatible structures adjusted to the specific peripheral nerve injury case can be manufactured.

Brain Regeneration Resembles Brain Cancer at Its Early Wound Healing Stage and Diverges From Cancer Later at Its Proliferation and Differentiation Stages.

Gliomas are the most frequent type of brain cancers and characterized by continuous proliferation, inflammation, angiogenesis, invasion and dedifferentiation, which are also among the initiator and sustaining factors of brain regeneration during restoration of tissue integrity and function. Thus, brain regeneration and brain cancer should share more molecular mechanisms at early stages of regeneration where cell proliferation dominates. However, the mechanisms could diverge later when the regenerative response terminates, while cancer cells sustain proliferation. To test this hypothesis, we exploited the adult zebrafish that, in contrast to the mammals, can efficiently regenerate the brain in response to injury. By comparing transcriptome profiles of the regenerating zebrafish telencephalon at its three different stages, i.e., 1 day post-lesion (dpl)-early wound healing stage, 3 dpl-early proliferative stage and 14 dpl-differentiation stage, to those of two brain cancers, i.e., low-grade glioma (LGG) and glioblastoma (GBM), we reveal the common and distinct molecular mechanisms of brain regeneration and brain cancer. While the transcriptomes of 1 dpl and 3 dpl harbor unique gene modules and gene expression profiles that are more divergent from the control, the transcriptome of 14 dpl converges to that of the control. Next, by functional analysis of the transcriptomes of brain regeneration stages to LGG and GBM, we reveal the common and distinct molecular pathways in regeneration and cancer. 1 dpl and LGG and GBM resemble with regard to signaling pathways related to metabolism and neurogenesis, while 3 dpl and LGG and GBM share pathways that control cell proliferation and differentiation. On the other hand, 14 dpl and LGG and GBM converge with respect to developmental and morphogenetic processes. Finally, our global comparison of gene expression profiles of three brain regeneration stages, LGG and GBM exhibit that 1 dpl is the most similar stage to LGG and GBM while 14 dpl is the most distant stage to both brain cancers. Therefore, early convergence and later divergence of brain regeneration and brain cancer constitutes a key starting point in comparative understanding of cellular and molecular events between the two phenomena and development of relevant targeted therapies for brain cancers.

Morphological and molecular features of early regeneration in the marine annelid Ophryotrocha xiamen

Regeneration capability varies in the phylum Annelida making them an excellent group to investigate the differences between closely related organisms. Several studies have described the process of regeneration, while the underlying molecular mechanism remains unclear, especially during the early stage (wound healing and blastema formation). In this study, the newly identified Ophryotrocha xiamen was used to explore the early regeneration. The detailed morphological and molecular analyses positioned O. xiamen within ‘labronica’ clade. We analyzed the morphological changes during regeneration process (0–3 days post amputation) and molecular changes during the early regeneration stage (1 day post amputation). Wound healing was achieved within one day and a blastema formed one day later. A total of 243 DEGs were mainly involved in metabolism and signal transduction. Currently known regeneration-related genes were identified in O. xiamen which could help with exploring the functions of genes involved in regeneration processes. According to their conserved motif, we identified 8 different Hox gene fragments and Hox5 and Lox2 were found to be absent in early regeneration and during regular growth. Our data can promote further use of O. xiamen which can be used as an experimental model for resolving crucial problems of developmental biology in marine invertebrates.

Pericyte‑derived extracellular vesicles‑mimetic nanovesicles improves peripheral nerve regeneration in mouse models of sciatic nerve transection.

Pericyte-derived extracellular vesicle-mimetic nanovesicles (PC-NVs) play an important role in the improvement of erectile function after cavernous nerve injury. However, the impact of PC-NVs on the peripheral nervous system (PNS), such as the sciatic nerve, is unclear. In this study, PC-NVs were isolated from mouse cavernous pericytes (MCPs). A sciatic nerve transection (SNT) model was established using 8-week-old C57BL/6J mice. The sciatic nerve was harvested 5 and 14 days for immunofluorescence and western blot studies. Function studies were evaluated by performing the rotarod test and walking track analysis. The results demonstrated that PC-NVs could stimulate endothelial cells, increase neuronal cell content, and increase macrophage and Schwann cell presence at the proximal stump rather than the distal stump in the SNT model, thereby improving angiogenesis and nerve regeneration in the early stage of sciatic nerve regeneration. In addition, PC-NVs also increased the expression of neurotrophic factors (brain-derived nerve growth factor, neurotrophin-3 and nerve growth factor) and the activity of the cell survival signaling pathway (PI3K/Akt signaling), and reduced the activity of the JNK signaling pathway. Additionally, after 8 weeks of local application of PC-NVs in SNT model mice, their motor and sensory functions were significantly improved, as assessed by performing the rotarod test and walking track analysis. In conclusion, the present study showed that the significant improvement of neurovascular regeneration in mice following treatment with PC-NVs may provide a favorable strategy for promoting motor and sensory regeneration and functional recovery of the PNS.

Multiple Ion Scaffold-Based Delivery Platform for Potential Application in Early Stages of Bone Regeneration.

Bone has the intrinsic capacity to regenerate itself, as long as the damage is small, through the sequential stimulation of specific phases, such as angiogenesis followed by osteogenesis. However, when the damage is extensive it is unable to regenerate and bone tissue engineering is used as an alternative. In this study, we developed a platform to allow the triple ion delivery with sequential delivery capacity to potentially stimulate antibacterial, angiogenic and osteogenic processes. The scaffold-based platform consisted of alginate/hydroxyapatite (HA) microparticles embedded in alginate fibers. Firstly, microparticles were developed using different ratios of alginate:HA using the spraying method, resulting in a high reproducibility of the technique. Microparticle size between 100–300 µm and ratio 1:40 resulted in a more spherical morphology and were selected for their incorporation into alginate fiber. Different amounts of copper and cobalt were added with the microparticles and alginate fiber, respectively, were used as model ions which could eventually modulate and mimic antimicrobial and angiogenic processes. Moreover, calcium ion was also incorporated in both, in order to provide the system with potential osteogenic properties together with HA. The multiple delivery of copper, cobalt and calcium released were in the therapeutic range as measured by induced coupled plasma (ICP), providing a promising delivery strategy for tissue engineering.

Head muscle fibro-adipogenic progenitors account for the tilted regeneration towards fibrosis

Branchiomeric head muscle is ontogenetically and phylogenetically distinct from somitic limb muscle, and they exhibit different regenerative capacity. Unique satellite cell property of head muscle could explain the impaired myofiber formation, but the underlying mechanism for fibrosis is still elusive. In this work, we first established a freezing-induced skeletal muscle regeneration model and made comparisons between the regeneration characteristics in tibialis anterior (TA) muscle and masseter (MAS) muscle. The process of myogenesis and fibrogenesis were investigated by histological, immunohistochemical and cellular analysis, to characterize the role of muscle satellite cell (MuSCs) and fibro-adipogenic progenitors (FAPs) in TA and MAS muscle regeneration. Our results revealed that FAPs infiltrated the fibrotic area during MAS muscle regeneration. In contrast to the rapid rise and fall of FAPs number at the early regeneration stages in TA muscle, the number of MAS FAPs increased to a plateau without descending till 14 days after injury. It is the first time that the pivotal role of FAPs in head muscle regeneration was characterized. The persistence of FAPs without timely clearance in the first two weeks of regeneration could be accountable for the head muscle fibrosis.

The effect of modifying the nanostructure of gelatin fiber scaffolds on early angiogenesis in vitro and in vivo

Early angiogenesis is one of the key challenges in tissue regeneration. Crosslinking mode and fiber diameter are critical factors to affect the adhesion and proliferation of cells. However, whether and how these two factors affect early angiogenesis remain largely unknown. To address the issue, the optimal crosslinking mode and fiber diameter of gelatin fiber membrane for early angiogenesis in vivo and in vitro were explored in this work. Compared with the post crosslinked gelatin fiber membrane with the same fiber diameter, the 700 nm diameter in situ crosslinked gelatin fiber membrane was found to have smaller roughness (230.67 ± 19 nm) and stronger hydrophilicity (54.77° ± 1.2°), which were suitable for cell growth and adhesion. Moreover, the in situ crosslinked gelatin fiber membrane with a fiber diameter of 1000 nm had significant advantages in early angiogenesis over the two with fiber diameters of 500 and 700 nm by up-regulating the expression of Ang1, VEGF, and integrin-β1. Our findings indicated that the in situ crosslinked gelatin fiber membrane with a diameter of 1000 nm might solve the problem of insufficient blood supply in the early stage of soft tissue regeneration and has broad clinical application prospects in promoting tissue regeneration.

Regeneration and Endogenous Phytohormone Responses to High-Temperature Stress Drive Recruitment Success in Hemiepiphytic Fig Species.

Exposure to high-temperature stress (HTS) during early regeneration in plants can profoundly shape seed germination, seedling growth, and development, thereby providing stress resilience. In this study, we assessed how the timing of HTS, which was implemented as 8 h in 40°C, could affect the early regeneration stages and phytohormone concentration of four hemiepiphytic (Hs) and four non-hemiepiphytic (NHs) Ficus species. Their seed germination, seedling emergence, and seedling survival probabilities and the concentrations of three endogenous phytohormones, abscisic acid (ABA), indole-3-acetic acid (IAA), and salicylic acid (SA) were assessed after HTS imposed during imbibition, germination, and emergence. In both groups, seeds were more sensitive to HTS in the early regeneration process; stress experienced during imbibition affected emergence and survival, and stress experienced during germination affected subsequent emergence. There was no effect from HTS when received after emergence. Survival was highest in hemiepiphytes regardless of the HTS treatment. The phytohormones showed growth form- and regeneration stage-specific responses to HTS. Due to the HTS treatment, both SA and ABA levels decreased in non-hemiepiphytes during imbibition and germination; during germination, IAA increased in hemiepiphytes but was reduced in non-hemiepiphytes. Due to the HTS treatment experienced during emergence ABA and IAA concentrations were greater for hemiepiphytes but an opposite effect was seen in the two growth forms for the SA concentration. Our study showed that the two growth forms have different strategies for regulating their growth and development in the early regeneration stages in order to respond to HTS. The ability to respond to HTS is an ecologically important functional trait that allows plant species to appropriately time their seed germination and seedling development. Flexibility in modulating species regeneration in response to HTS in these subtropical and tropical Ficus species could provide greater community resilience under climate change.

Regenerative Polarity of the Fin Ray in Zebrafish Caudal Fin and Related Tissue Formation on the Cut Surface.

Zebrafish caudal fin rays are used as a model system for regeneration because of their high regenerative ability, but studies on the regeneration polarity of the fin ray are limited. To investigate this regeneration polarity, we made a hole to excise part of the fin ray and analyzed the regeneration process. We confirmed that the fin rays always regenerated from the proximal margin toward the distal margin, as previously reported; however, regeneration-related genes were expressed at both the proximal and distal edges of the hole in the early stage of regeneration, suggesting that the regenerative response also occurs at the distal edge. One difference between the proximal and distal margins is a sheet-like tissue that is formed on the apical side of the regenerated tissue at the proximal margin. This sheet-like tissue was not observed at the distal edge. To investigate whether the distal margin was also capable of forming this sheet-like tissue and subsequent regeneration, we kept the distal margin separated from the proximal margin by manipulation. Consequently, the sheet-like tissue was formed at the distal margin and regeneration of the fin ray was also induced. The regenerated fin rays from the distal margin protruded laterally from the caudal fin and then bent distally, and their ends showed the same characteristics as those of the normal fin rays. These results suggest that fin rays have an ability to regenerate in both directions; however, under normal conditions, regeneration is restricted to the proximal margin because the sheet-like tissue is preferentially formed on the apical side of the regenerating tissue from the proximal margin.

Short-term exposure to perfluorotetradecanoic acid affects the late-stage regeneration of Leydig cells in adult male rats

Perfluorotetradecanoic acid (PFTeDA) is one of perfluoroalkyl substances widely found in the environment. PFTeDA may cause the dysfunction of male reproductive system. However, whether PFTeDA affects the regeneration of Leydig cells remains unclear. The objective of this study was to examine the effects of short-term exposure of PFTeDA on the late-stage maturation of Leydig cells. Fifty-four adult Sprague-Dawley male rats were daily gavaged with PFTeDA (0, 10, or 20 mg/kg body weight) for 10 days, and then were injected intraperitoneally with ethylene dimethane sulfonate (EDS, 75 mg/kg body weight/once) to ablate Leydig cells to induce their regeneration. On day 21 (early stage) and 56 (late stage) after EDS, hormone levels, gene expression, and protein levels were measured. PFTeDA did not affect the early stage of Leydig cell regeneration, because it had no effect on serum testosterone, luteinizing hormone, and follicle-stimulating hormone levels, Leydig cell number, and its gene and protein expression. PFTeDA significantly reduced serum testosterone level and down-regulated the expression of Leydig cell genes (Cyp11a1, Hsd3b1, Cyp17a1, Hsd17b3, Hsd11b1, and Insl3) and their proteins (CYP11A1, HSD3B1, CYP17A1, HSD17B3, and INSL3), decreased the phosphorylation of AKT1 and ERK1/2, as well as lowered sperm count in the epididymis at 20 mg/kg. In conclusion, short-term exposure to PFTeDA blocks the late-stage maturation of Leydig cells.

Arabidopsis ATXR2 represses de novo shoot organogenesis in the transition from callus to shoot formation.

Plants exhibit high regenerative capacity, which is controlled by various genetic factors. Here, we report that ARABIDOPSIS TRITHORAX-RELATED 2 (ATXR2) controls de novo shoot organogenesis by regulating auxin-cytokinin interaction. The auxin-inducible ATXR2 Trithorax Group (TrxG) protein temporally interacts with the cytokinin-responsive type-B ARABIDOPSIS RESPONSE REGULATOR 1 (ARR1) at early stages of shoot regeneration. The ATXR2-ARR1 complex binds to and deposits the H3K36me3 mark in the promoters of a subset of type-A ARR genes, ARR5 and ARR7, thus activating their expression. Consequently, the ATXR2/ARR1-type-A ARR module transiently represses cytokinin signaling and thereby de novo shoot regeneration. The atxr2-1 mutant calli exhibit enhanced shoot regeneration with low expression of ARR5 and ARR7, which ultimately upregulates WUSCHEL (WUS) expression. Thus, ATXR2 regulates cytokinin signaling and prevents premature WUS activation to ensure proper cell fate transition, and the auxin-cytokinin interaction underlies the initial specification of shoot meristem in callus.

Association between native tree sapling and spiny shrub mitigates browsing damage produced by large herbivores in fire-degraded forests

Spiny shrubs can generate positive interactions with palatable trees in sites affected by abiotic and biotic stress (e.g. fire, and consumption by herbivores). The objective of this study was to assess the role of native shrubs (Berberis microphylla) in mitigating large herbivore browsing on tree saplings (Nothofagus antarctica) in degraded (burnt) forests of the central area of Tierra del Fuego, Argentina. For that, we assessed, in 16 forest sites, the browsing intensity and the browsing height in N. antarctica saplings that were alone (n = 10 per site), and in saplings that were spatially associated with B. microphylla shrubs (n = 10 per site). The tree and shrub height were also measured. The results indicated that browsing damage on ñire trees occurred at a higher height and with lower intensity in the tree-shrub association than in trees that were alone. Furthermore, the browsing height was positively related with the shrub height and with the tree height. The shorter trees with greater shrub cover were more protected. Our results indicate that the presence of spiny shrubs decreases the damage caused by browsing, and protects trees during the early regeneration stage. However, the structural characteristics of shrubs are determinant of the protecting effect in these Patagonian forests. This tree-shrub interaction is highly relevant in the dynamics of degraded forests to ensure the system continuity in ecological and productivity terms.

Structural changes to forests during regeneration affect water flux partitioning, water ages and hydrological connectivity: Insights from tracer-aided ecohydrological modelling

Abstract. Increasing rates of biodiversity loss are adding momentum to efforts seeking to restore or rewild degraded landscapes. Here, we investigated the effects of natural forest regeneration on water flux partitioning, water ages and hydrological connectivity, using the tracer-aided ecohydrological model EcH2O-iso. The model was calibrated using ∼ 3.5 years of diverse ecohydrological and isotope data available for a catchment in the Scottish Highlands, an area where impetus for native pinewood regeneration is growing. We then simulated two land cover change scenarios that incorporated forests at early (dense thicket) and late (old open forest) stages of regeneration, respectively. Changes to forest structure (proportional vegetation cover, vegetation heights and leaf area index of pine trees) were modelled for each stage. The scenarios were then compared to a present-day baseline simulation. Establishment of thicket forest had substantial ecohydrological consequences for the catchment. Specifically, increased losses to transpiration and, in particular, interception evaporation drove reductions in below-canopy fluxes (soil evaporation, groundwater (GW) recharge and streamflow) and generally slower rates of water turnover. The greatest reductions in streamflow and connectivity were simulated for summer baseflows and small to moderate events during summer and the autumn/winter rewetting period. This resulted from the effect of local changes to flux partitioning in regenerating areas on the hillslopes extending to the wider catchment by reducing downslope GW subsidies that help sustain summer baseflows and saturation in the valley bottom. Meanwhile, higher flows were relatively less affected, especially in winter. Despite the generally drier state of the catchment, simulated water ages suggested that the increased transpiration demands of the thicket forest could be satisfied by moisture carried over from previous seasons. The more open nature of the old forest generally resulted in water fluxes, water ages and connectivity returning towards baseline conditions. Our work implies that the ecohydrological consequences of natural forest regeneration depend on the structural characteristics of the forest at different stages of development. Consequently, future land cover change investigations need to move beyond consideration of simple forest vs. non-forest scenarios to inform sustainable landscape restoration efforts.

Altitude and landslide scale regulated the assembly of grassland communities on landslides during the recovery process after the magnitude 8.0 Wenchuan earthquake, China

Earthquakes are one of the worst natural disasters, causing environmental fluctuations and ecosystem degradation. Previous studies have evaluated the macro-characteristics of vegetation recovery after earthquakes. However, the internal mechanisms driving community assembly and diversity remain poorly understood. This study was located in Longxi-Hongkou National Natural Reserve, China. We investigated the current vegetation status of typical large-scale landslides in regions with the highest intensity of earthquake damage in the area affected by the 2008 magnitude 8.0 earthquake, and studied the community assembly using phylogenetic methods. Relationships between community assembly and influencing factors were carried out using generalized linear models (GLMs) and redundancy analysis (RDA). Our findings suggested that the communities in areas of larger landslides were dominated by herbs 11 years after the earthquake and had not yet regenerated to the original community. The average net relatedness index (NRI) and nearest taxon index (NTI) of the communities were 0.293 and 0.482, respectively, which were higher than those of the null model. These results showed a clustered phylogenetic pattern and suggested that the community assembly process in the early regeneration stage after the earthquake supported the environmental filtering hypothesis. Altitude negatively impacted species diversity, NRI, NTI and phylogenetic diversity. This result suggested that the community assembly process was more dependent on abiotic regulation than biotic regulation at high altitudes after earthquakes. Species diversity, NRI, NTI and phylogenetic diversity were also impacted by landslide intensity; however, the responses to landslide intensity were not completely equivalent for landslides of varying widths and lengths. Slope aspect impacted the phylogenetic diversity of the regenerating community after the earthquake; that is, the sunny slopes had higher phylogenetic diversity than the shady slopes. Soil type, soil organic carbon, soil nitrogen, soil moisture, and soil phosphorous impacted community assembly. Collectively, This work showed evidence supporting the abiotic regulation of community assembly after earthquakes and reported the key abiotic influencing factors. It is important for a general understanding of community assembly after strong disturbances in temperate forests and beyond and can also provide a theoretical basis for ecological restoration engineering design.

Resorbable Mg2+-Containing Phosphates for Bone Tissue Repair.

Materials based on Mg2+-containing phosphates are gaining great relevance in the field of bone tissue repair via regenerative medicine methods. Magnesium ions, together with condensed phosphate ions, play substantial roles in the process of bone remodeling, affecting the early stage of bone regeneration through active participation in the process of osteosynthesis. In this paper we provide a comprehensive overview of the usage of biomaterials based on magnesium phosphate and magnesium calcium phosphate in bone reconstruction. We consider the role of magnesium ions in angiogenesis, which is an important process associated with osteogenesis. Finally, we summarize the biological properties of calcium magnesium phosphates for regeneration of bone.

Mineral Trioxide Aggregate (MTA) Upregulates the Expression of DMP1 in Direct Pulp Capping in the Rat Molar.

Mineral trioxide aggregate (MTA) is an alternative endodontic material that predicts conductive or inductive calcified tissue formation from immature pulp mesenchymal stem cells (IPMSCs). The purpose of this study was to investigate whether MTA could promote reparative odontoblast differentiation via IPMSCs in the early phase of regeneration and compare with calcium hydroxide (CH). Direct pulp capping using calcium hydroxide (CH), MTA, and MTA with platelet-rich plasma (MTA + PRP) was performed on maxillary first molars of 8-week-old male Wistar rats (n = 36). After 3, 7, or 14 days, the teeth were analyzed for mineral density (MD) and volume of MD (VMD) via micro-focusing computed tomography (µCT), nestin, dentin matrix acidic phosphoprotein 1 (DMP1) immunohistochemistry, and real-time PCR for DMP1 mRNA expression. MTA stimulated the early phase differentiation of the IPMSCs into odontoblasts, with positive results for nestin and DMP1 compared with CH. Moreover, MTA + PRP stimulated calcified granule and dentin bridge formation through calcium mineral deposition, following the induction of DMP1 mRNA expression in IPMSCs. Our results suggested that the combination of MTA and PRP is an effective and clinically applicable method for activating endogenous dental pulp stem cells into odontoblasts in the early stages of pulp regeneration.

Hybrid material mimics a hypoxic environment to promote regeneration of peripheral nerves

Between nerve defects, a bridge formed by multiple cells is the fundamental structure for guiding axons across this damaged region. Here, we developed a functional material that mimics hypoxia during the early stages of nerve regeneration by deferoxamine. We used this material and single-cell sequencing to analyze the “bridge” structure between peripheral nerve defects. We found that hypoxia in damaged tissues might play a key role in stimulating macrophages, promoting endothelial-to-mesenchymal transition, and driving the migration of endothelial cells to the injured region to form regenerative bridge tissue and guide the subsequent regeneration of Schwann cells and axons. The results showed that the final nerve defect repair outcomes were similar with autografts after intervention by this material. This study challenges the view that hypoxia is exclusively involved in peripheral nerve regeneration and provides a potentially valuable candidate material for clinical use.

Met and Cxcr4 cooperate to protect skeletal muscle stem cells against inflammation-induced damage during regeneration.

Acute skeletal muscle injury is followed by an inflammatory response, removal of damaged tissue, and the generation of new muscle fibers by resident muscle stem cells, a process well characterized in murine injury models. Inflammatory cells are needed to remove the debris at the site of injury and provide signals that are beneficial for repair. However, they also release chemokines, reactive oxygen species, as well as enzymes for clearance of damaged cells and fibers, which muscle stem cells have to withstand in order to regenerate the muscle. We show here that MET and CXCR4 cooperate to protect muscle stem cells against the adverse environment encountered during muscle repair. This powerful cyto-protective role was revealed by the genetic ablation of Met and Cxcr4 in muscle stem cells of mice, which resulted in severe apoptosis during early stages of regeneration. TNFα neutralizing antibodies rescued the apoptosis, indicating that TNFα provides crucial cell-death signals during muscle repair that are counteracted by MET and CXCR4. We conclude that muscle stem cells require MET and CXCR4 to protect them against the harsh inflammatory environment encountered in an acute muscle injury.

Seedling emergence and environmental filters determine Ficus recruitment in a subtropical landscape

Early regeneration, involving the transition from a seed into a successfully recruited seedling, is a complex process characterized by high mortality rates and the strong association with environmental filters. However, we have limited understanding of the factors and transitional regeneration stages that lead to these broad mortality events, even in species that are key for ecosystem function and comprise the bulk of species in diversity assessments, such as Ficus species. Using eight, 1-ha long term monitoring plots, established along an elevation gradient in tropical to subtropical forests in southern China, and partitioning the early plant regeneration process into four stages (seed germination, seedling emergence, establishment and recruitment), we conducted a seed addition experiment to assess the factors driving the early regeneration process. We used four hemi-epiphytic and four non-hemiepiphytic Ficus species, specifically asking whether elevation, microsite conditions and growth form affect the success of regeneration outcomes. We observed that Ficus early regeneration was highly limited throughout the subtropical landscape with similar seed germination probabilities along all elevations, but greater seedling emergence at higher elevations. Growth form and elevation interacted significantly during seed germination and seedling emergence stages. The strongest filtering effect was observed during seedling emergence, with greater canopy openness facilitating, but drier and cooler soils being less conducive to seedling emergence, especially for hemiepiphytic species. Once emerged, Ficus seedlings were released from the environmental filters assessed here, yet, continued to face successively greater mortality during establishment and recruitment. Greater seed germination and seedling emergence on the ground observed in hemiepiphytes indicate that they may benefit from evolutionary traits associated with canopy regeneration under water and temperature stress, but only at the early regeneration stages, as seedling establishment and recruitment were similar in both groups. In general, our results demonstrate that even when seeds are available, successful recruitment is strongly limited by microsite conditions and growth form, through filtering mechanisms associated with the transition from seedling emergence to seedling establishment. The identification of environmental filters that operate during the complex and vulnerable early regeneration stages provides a baseline to develop management strategies for ex situ conservation, as well as in situ population reestablishment for restoration.