Tissue self-repair after trauma indicated by immunohistochemical staining and semi-quantitative analysis of PCNA, Col I and OCN.

<p>(A, C, E) Immunohistochemical assays of the expression of PCNA (A), Col I (C) and OCN (E). (B, D, F) Bar graphs represent their expression density in unit area of bone marrow or unit area of bone trabeculae. (A) OBs; (C) a. OBs, b. osteocytes, c. trabeculae, d. VECs, e. OCs. The expression of PCNA decreased after trauma, and began to be detected in OBs 2 weeks after trauma and was maintained during the following time. Col I increased in the bone marrow for less than 1 week after trauma, and was significantly accumulated in the trabeculae by the end of the study. Both OBs and OCs also expressed increased levels of Col I following trauma. A significant decrease in OCN was detected 3 days after trauma, but it increased significantly 2 weeks later. High levels of OCN were consistently associated with peri-VECs and the fibrous medulla. N: normal group; 3 d, 1 w, 2 w, 3 w: 3 days, 1 week, 1 weeks, 3 weeks post-trauma, respectively. Quantification was based on at least 10 fields per section. *<i>P</i><0.05 vs. normal group.</p

Histopathological images of the traumatic ONFH rabbit model, and immunohistochemical staining and semi-quantitative analysis of vWF and CD105.

<p>(A) Representative images of a section of femoral head including cartilage stained with H&E. Scale bar = 200 µm. In the injured femoral head, the number of empty lacunae increased and hematopoietic tissue diminished significantly, but cartilage did not exhibit any obvious pathological changes. Immature fibrotic tissue and appositional bone formation were observed under the cartilage from 3 days after trauma, followed by an increase in the number of OBs 2 weeks later. a. empty lacuna; b. immature fibrotic tissue; c. appositional bone formation; d. OBs. (B) Bar graphs represent the ratio of bone marrow cells to the area of bone marrow (a), the ratio of empty lacunae to the area of trabeculae (b), thinning trabeculae (c) and grey scale (d), respectively. (C) Immunohistochemical assays of vWF expression. Scale bar = 50 µm. (D) Blood vessels were counted according to positive staining of vWF in combination with appropriate vessel structure. Following trauma, the structure of the blood vessels became increasingly compromised and the number of blood vessels decreased. Arrows = microvessels or arteries. (E) Immunohistochemical assays of vWF expression. Scale bar = 20 µm. (F) Bar graphs represent the expression density of CD105 as unit area of bone marrow or unit area of bone trabeculae. The expression of CD105 until 3 weeks after trauma suggested the presence of revascularization in the injured femoral head. N: normal group; 3 d, 1 w, 2 w, 3 w: 3 days, 1 week, 2 weeks, 3 weeks post-trauma, respectively. Quantification was based on at least 10 fields per section. *<i>P</i><0.05 vs. normal group.</p

Immunohistochemical staining and semi-quantitative analysis of VEGF and Col I.

<p>(A, C) Immunohistochemical assays of the expression of VEGF (A) and Col I (C). (B, D) Bar graphs represent the expression density of VEGF and Col I as unit area of bone marrow or unit area of bone trabeculae. Transplantation of MSCs reversed the decline of VEGF and Col I expression in the marrow, but decreased the accumulation of Col I in the trabeculae. The most significant recovery was seen in the ONFH+MSC+HGF group. Quantification was based on at least 10 fields per section. *<i>P</i><0.05 vs. normal group.</p

Identification of MSC pluripotent potential.

<p>Undifferentiated MSCs (a–e), osteoblasts (f–j), chondroblasts (k–o), and adipocytes (p–t) were left unstained (a, f, k, & p), or stained with Alizarin Red at day 21 (b, g, l, & q), NBT-BCIP at day 14 (c, h, m, & r), Alcian blue at day 27 (d, i, n, & s), or Oil Red O at day 27 (e, j, o, & t). Scale bar = 50 µm.</p

Immunohistochemical detection and semi-quantitative analysis of HGF expression (A), phosphorylation of ERK1/2 (p-ERK1/2) (B) and Akt (p-Akt) (C).

<p>There was a little increase in HGF after trauma. After the transplantation of MSCs, the HGF level increased significantly at as early as 2 days, which was concomitant with increased p-ERK1/2. The HGF level decreased gradually for 2 weeks after transplantation, followed by a significant increase in Akt activation. The effects were most marked in the animals treated with HGF-transgenic MSCs. ^*<i>P</i><0.05, compared with the normal group. ^#<i>P</i><0.05, compared with the non-infected MSC-treated group. Scale bar = 50 µm.</p

Histopathological examination of treatment efficacy in traumatic ONFH by H&E staining, and immunohistochemical staining and semi-quantitative analysis of CD105.

<p>(A) Representative image of a section of femoral head stained with H&E. Scale bar = 200 µm. In the treated femoral head, the number of empty lacunae decreased and hematopoietic tissue partially recovered, accompanied by an increase in the number of OBs, which was most significant in animals that received HGF-transgenic MSCs. a. empty lacuna; b. OBs. (B) Bar graph (left panel) represents the ratio of bone marrow cells to the area of bone marrow. A second bar graph (right panel) represents the ratio of empty lacunae to the area of trabeculae. (C) Immunohistochemical assays of CD105 expression. Scale bar = 20 µm. (D) Bar graphs represent the expression density of CD105 as unit area of bone marrow or unit area of bone trabeculae. After MSC transplantation, the expression of CD105 increased compared with that of untreated ONFH group, indicating the occurrence of revascularization. Quantification was based on at least 10 fields per section. *<i>P</i><0.05.</p

Conventional CT and MR examination of the traumatic ONFH rabbit model established in this study.

<p>Left column: Conventional CT examination (coronal reconstruction). With progression of the disease, the density of cancellous bone became more heterogeneous and eventually lost its symmetry. Arrows indicate the femoral head. Middle column: MR-FS T2-weighted imaging (coronal plane) detected a signal that became increasingly uneven for up to 2 weeks after trauma. A signal with high intensity was observed at the fracture line, suggesting edema was present in the bone marrow. Right column: MR T1-weighted imaging (coronal plane) detected a loss of normal signal within the femoral head. Circles indicate the femoral head. N: normal group; 3 d, 1 w, 2 w, 3 w: 3 days, 1 week, 2 weeks, 3 weeks post-trauma, respectively.</p

Nanosensor-Driven Detection of Neuron-Derived Exosomal Aβ₄₂ with Graphene Electrolyte-Gated Transistor for Alzheimer’s Disease Diagnosis

Blood-based tests have sparked tremendous attention in non-invasive early diagnosis of Alzheimer’s disease (AD), a most prevalent neurodegenerative malady worldwide. Despite significant progress in the methodologies for detecting AD core biomarkers such as Aβ42 from serum/plasma, there remains cautious optimism going forward due to its controversial diagnostic value and disease relevance. Here, a graphene electrolyte-gated transistor biosensor is reported for the detection of serum neuron-derived exosomal Aβ42 (NDE-Aβ42), which is an emerging, compelling trove of blood biomarker for AD. Assisted by the antifouling strategy with the dual-blocking process, the noise against complex biological background was considerably reduced, forging an impressive sensitivity gain with a limit of detection of 447 ag/mL. An accurate detection of SH-SY5Y-derived exosomal Aβ42 was also achieved with highly conformable enzyme-linked immunosorbent assay results. Importantly, the clinical analysis for 27 subjects revealed the immense diagnostic value of NDE-Aβ42, which can outclass that of serum Aβ42. The developed electronic assay demonstrates, for the first time, nanosensor-driven NDE-Aβ42 detection, which enables a reliable discrimination of AD patients from non-AD individuals and even the differential diagnosis between AD and vascular dementia patients, with an accuracy of 100% and a Youden index of 1. This NDE-Aβ42 biosensor defines a robust approach for blood-based confident AD ascertain

sj-docx-1-cll-10.1177_09636897231210750 – Supplemental material for HIF-1α-Induced Mitophagy Regulates the Regenerative Outcomes of Stem Cells in Fat Transplantation

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