1.A prospective study on application of human umbilical cord mesenchymal stem cells combined with autologous Meek microskin transplantation in patients with extensive burns.
Tian Tian YAN ; Rong XIAO ; Ying WANG ; Guo An LIN ; Yin ZHENG ; Hui ZHAO ; Wen Jun LI ; Xin Zhi SHANG ; Jin Song MENG ; Dong Sheng HU ; Song LI ; Chao WANG ; Zhi Chen LIN ; Hong Chang CHEN ; Dong Yan ZHAO ; Di TANG
Chinese Journal of Burns 2023;39(2):114-121
Objective: To investigate the effects of human umbilical cord mesenchymal stem cells (hUCMSCs) combined with autologous Meek microskin transplantation on patients with extensive burns. Methods: The prospective self-controlled study was conducted. From May 2019 to June 2022, 16 patients with extensive burns admitted to the 990th Hospital of PLA Joint Logistics Support Force met the inclusion criteria, while 3 patients were excluded according to the exclusion criteria, and 13 patients were finally selected, including 10 males and 3 females, aged 24-61 (42±13) years. A total of 20 trial areas (40 wounds, with area of 10 cm×10 cm in each wound) were selected. Two adjacent wounds in each trial area were divided into hUCMSC+gel group applied with hyaluronic acid gel containing hUCMSCs and gel only group applied with hyaluronic acid gel only according to the random number table, with 20 wounds in each group. Afterwards the wounds in two groups were transplanted with autologous Meek microskin grafts with an extension ratio of 1∶6. In 2, 3, and 4 weeks post operation, the wound healing was observed, the wound healing rate was calculated, and the wound healing time was recorded. The specimen of wound secretion was collected for microorganism culture if there was purulent secretion on the wound post operation. In 3, 6, and 12 months post operation, the scar hyperplasia in wound was assessed using the Vancouver scar scale (VSS). In 3 months post operation, the wound tissue was collected for hematoxylin-eosin (HE) staining to observe the morphological changes and for immunohistochemical staining to observe the positive expressions of Ki67 and vimentin and to count the number of positive cells. Data were statistically analyzed with paired samples <i>ti> test and Bonferronni correction. Results: In 2, 3, and 4 weeks post operation, the wound healing rates in hUCMSC+gel group were (80±11)%, (84±12)%, and (92±9)%, respectively, which were significantly higher than (67±18)%, (74±21)%, and (84±16)% in gel only group (with <i>ti> values of 4.01, 3.52, and 3.66, respectively, <i>Pi><0.05). The wound healing time in hUCMSC+gel group was (31±11) d, which was significantly shorter than (36±13) d in gel only group (<i>ti>=-3.68, <i>Pi><0.05). The microbiological culture of the postoperative wound secretion specimens from the adjacent wounds in 2 groups was identical, with negative results in 4 trial areas and positive results in 16 trial areas. In 3, 6, and 12 months post operation, the VSS scores of wounds in gel only group were 7.8±1.9, 6.7±2.1, and 5.4±1.6, which were significantly higher than 6.8±1.8, 5.6±1.6, and 4.0±1.4 in hUCMSC+gel group, respectively (with <i>ti> values of -4.79, -4.37, and -5.47, respectively, <i>Pi><0.05). In 3 months post operation, HE staining showed an increase in epidermal layer thickness and epidermal crest in wound in hUCMSC+gel group compared with those in gel only group, and immunohistochemical staining showed a significant increase in the number of Ki67 positive cells in wound in hUCMSC+gel group compared with those in gel only group (<i>ti>=4.39, <i>Pi><0.05), with no statistically significant difference in the number of vimentin positive cells in wound between the 2 groups (<i>Pi>>0.05). Conclusions: The application of hyaluronic acid gel containing hUCMSCs to the wound is simple to perform and is therefore a preferable route. Topical application of hUCMSCs can promote healing of the autologous Meek microskin grafted area in patients with extensive burns, shorten wound healing time, and alleviate scar hyperplasia. The above effects may be related to the increased epidermal thickness and epidermal crest, and active cell proliferation.
Female
;
Humans
;
Male
;
Burns/surgery*
;
Cicatrix
;
Eosine Yellowish-(YS)
;
Hyaluronic Acid/therapeutic use*
;
Hyperplasia
;
Ki-67 Antigen
;
Prospective Studies
;
Umbilical Cord
;
Vimentin
;
Young Adult
;
Adult
;
Middle Aged
2.The effect of fluorescein angiography on renal function: A meta-analysis and systematic review
Kevin Elissandro C. Gumabon ; Paolo Nikolai H. So ; Anne Margaret J. Ang
Acta Medica Philippina 2023;57(3):42-49
Background:
Contrast-induced nephropathy (CIN) is a complication that occurs in patients undergoing an imaging procedure with intravenous injection of contrast media, most notably iodinated dyes. Fluorescein angiography is a diagnostic procedure performed by ophthalmologists to determine abnormalities in retinal blood vessels. It uses sodium fluorescein, an organic dye, to capture and visualize these blood vessels. There have been conflicting data and practices on how to approach the procedure especially in patients with renal insufficiency.
Objective:
To determine the risk of CIN among patients undergoing fluorescein angiography.
Methods:
We searched PubMed, HerdIn, Cochrane Library, and Google Scholar, for published articles on the
topic. Other sources were searched for unpublished data or ongoing clinical trials. All research articles pertaining to fluorescein angiography and its effect on renal function with serum creatinine monitoring were included. Two independent authors separately screened records, assessed full texts, and extracted data. We used RevMan computer software to analyze data from the included studies. The primary outcome was the risk of CIN among patients undergoing fluorescein angiography based on the differences on serum creatinine levels and estimated glomerular filtration rates pre- and post-angiography, while the secondary outcome included risk factors for CIN.
Results:
A total of 6 studies were included in the meta-analysis. Four studies had poor quality as assessed using the Newcastle-Ottawa Scale. One study was deemed to have good quality. Data analysis showed that hemoglobin (p = 0.002) and albumin (p < 0.001) levels may be associated with CIN using sodium fluorescein but were not independent risk factors for CIN (multivariable logistic regression, p = 0.648 and p = 0.069, respectively); while sex, diabetes mellitus and chronic kidney disease were not significantly associated. As a primary outcome, only 6.8% of included patients had CIN with serum creatinine levels post-exposure showed significant differences from baseline values (mean difference 0.05; 95% CI 0.02, 0.07; I2 = 49%), but translating it to eGFR yielded non-significant differences (mean difference -0.37; 95% CI -2.33, 1.59; I2 = 0%).
Conclusion
Among patients undergoing fluorescein angiography, sodium fluorescein does not pose an increased risk for CIN.
fluorescein angiography
;
renal function
3.Acupuncture for aqueous deficiency dry eye: a randomized controlled trial.
Xue LIU ; Wei-Ping GAO ; Nan ZHAO ; Rui-Ping JIE ; Cheng-Yong LIU
Chinese Acupuncture & Moxibustion 2023;43(11):1235-1238
OBJECTIVES:
To compare the clinical efficacy and safety of acupuncture and sodium hyaluronate eye drop in the treatment of aqueous deficiency dry eye.
METHODS:
A total of 60 patients (120 eyes) with aqueous deficiency dry eye were randomly divided into an observation group (30 cases, 1 case dropped out) and a control group (30 cases, 1 case dropped out). In the control group, sodium hyaluronate eye drop were used, one drop at a time, 4 times a day, for 14 consecutive days. In the observation group, acupuncture was applied at bilateral Shangjingming (Extra), Cuanzhu (BL 2), Sizhukong (TE 23), Taiyang (EX-HN 5), and Tongziliao (GB 1) , once a day, treatment for 6 days with the interval of 1 day was required, for 14 consecutive days. The tear meniscus height (TMH), Schirmer Ⅰ test (SⅠT), ocular surface disease index (OSDI) score, non-invasive tear break-up time (NIBUT), and corneal fluorescein sodium staining (FLS) score were compared between the two groups before and after treatment, and the safety of the treatment of the two groups was observed.
RESULTS:
Compared with those before treatment, after treatment, TMH, SⅠT and NIBUT were increased (<i>Pi><0.01, <i>Pi><0.05), and FLS scores were decreased (<i>Pi><0.01) in the two groups; the score of OSDI was reduced (<i>Pi><0.01) in the observation group. After treatment, in the observation group, TMH and SⅠT were higher than those in the control group (<i>Pi><0.01), and the score of OSDI was lower than that in the control group (<i>Pi><0.01). No adverse reactions and adverse events were observed in the two groups.
CONCLUSIONS
Acupuncture and sodium hyaluronate eye drop can both effectively treat aqueous deficiency dry eye, acupuncture has obvious advantages in improving TMH and basic tear secretion, and reducing the subjective symptoms of patients. Acupuncture for dry eye is safe.
Humans
;
Hyaluronic Acid
;
Acupuncture Therapy
;
Dry Eye Syndromes/therapy*
;
Eye
;
Tears
;
Ophthalmic Solutions
;
Fluorescein
4.The Effect of Intense Pulsed Light Treatment for Chronic Hordeolum.
Ke YANG ; Ya WEN ; Lei ZHU ; Jia Yu BAO ; Shang LI ; Ying Hui WANG ; Jun FENG ; Lei TIAN ; Ying JIE
Biomedical and Environmental Sciences 2023;36(11):1005-1014
OBJECTIVE:
To evaluate the effect of intense pulsed light (IPL) in the treatment of chronic hordeolum.
METHODS:
Patients with chronic hordeolum who underwent IPL treatment were enrolled in this study. According to the severity of hordeolum, the patients were treated with IPL 3 to 5 times. Patients' satisfaction and visual analog scale scores for ocular discomfort symptoms before and after treatment were collected. The number, congestion, long diameter, short diameter and area of nodules were also recorded and measured. Finally, eyelid margin signs, meibum quality, meibomian gland expressibility, meibomian gland dropout, tear meniscus height, and corneal fluorescein staining were scored.
RESULTS:
20 patients were enrolled in this study. The eyelid margins were congestive and swollen, with blunt rounding or irregularity. The meibum was cloudy or toothpaste-like. The meibomian gland expressibility, meibomian gland dropout and tear meniscus height were reduced. The cornea showed scattered fluorescein staining. After treatment, score of visual analog scale, congestion and size of nodules were significantly reduced. Eyelid margin signs, meibum quality, meibomian gland expressibility, tear meniscus height and corneal fluorescein staining scores were improved. Meibomian gland dropout had no significant change. No side effects occurred during treatment.
CONCLUSIONS
IPL is beneficial for the treatment of chronic hordeolum.
Humans
;
Hordeolum
;
Meibomian Glands
;
Tears
;
Fluoresceins
5.Chemical constituents from stems and leaves of Cratoxylum cochinchinense and their inhibitory effects on proliferation of synoviocytes in vitro.
Yong ZHANG ; Ni-Fei SHI ; Zhen XIE ; Yi-Meng ZHAO ; Cai-Huan LIANG ; Ya-Yuan DENG ; Ran WANG ; Yan-Ping LIU ; Yan-Hui FU
China Journal of Chinese Materia Medica 2023;48(18):5014-5023
The chemical constituents from the stems and leaves of Cratoxylum cochinchinense were isolated and purified using silica gel, ODS gel, and Sephadex LH-20 gel column chromatography, as well as preparative HPLC. The chemical structures of all isolated compounds were identified on the basis of their physicochemical properties, spectroscopic analyses, and the comparison of their physicochemical and spectroscopic data with the reported data in literature. As a result, 21 compounds were isolated from the 90% ethanol extract of the stems and leaves of C. cochinchinense, which were identified as cratocochine(1), 1-hydroxy-3,7-dimethoxyxanthone(2), 1-hydroxy-5,6,7-trimethoxyxanthone(3), ferrxanthone(4), 3,6-dihydroxy-1,5-dimethoxyxanthone(5), 3,6-dihydroxy-1,7-dimethoxyxanthone(6), 1,2,5-trihydroxy-6,8-dimethoxyxanthone(7), securixanthone G(8), gentisein(9), 3,7-dihydroxy-1-methoxyxanthone(10), pancixanthone B(11), garcimangosxanthone A(12), pruniflorone L(13), 9-hydroxy alabaxanthone(14), cochinchinone A(15), luteolin(16), 3,5'-dimethoxy-4',7-epoxy-8,3'-neolignane-5,9,9'-triol(17), N-benzyl-9-oxo-10E,12E-octadecadienamide(18), 15-hydroxy-7,13E-labdadiene(19), stigmasta-4,22-dien-3-one(20), and stigmast-5-en-3β-ol(21). Among these isolates, compound 1 was a new xanthone, compounds 2-5, 7, 8, 12, and 16-21 were isolated from the Cratoxylum plant for the first time, and compounds 11 and 13 were obtained from C. cochinchinense for the first time. Furthermore, all isolated compounds 1-21 were appraised for their anti-rheumatoid arthritis activities by MTS method through measuring their anti-proliferative effect on synoviocytes in vitro. As a result, xanthones 1-15 displayed notable anti-rheumatoid arthritis activities, which showed inhibitory effects on the proliferation of MH7A synoviocytes with the IC_(50) values ranging from(8.98±0.12) to(228.68±0.32) μmol·L~(-1).
Synoviocytes
;
Clusiaceae/chemistry*
;
Xanthones/analysis*
;
Plant Leaves/chemistry*
;
Cell Proliferation
;
Arthritis
6.A new xanthone from hulls of Garcinia mangostana and its cytotoxic activity.
Feng-Ning ZHAO ; Qi NIU ; Die XIAO ; Hao-Nan XU ; Hao-Xin WANG ; Rong-Lu BI ; Hong-Ping HE ; Zhi-Yong JIANG
China Journal of Chinese Materia Medica 2023;48(21):5817-5821
Eight compounds were isolated from ethyl acetate fraction of 80% ethanol extract of the hulls of Garcinia mangostana by silica gel, Sephadex LH-20 column chromatography, as well as prep-HPLC methods. By HR-ESI-MS, MS, 1D and 2D NMR spectral analyses, the structures of the eight compounds were identified as 16-en mangostenone E(1), α-mangostin(2), 1,7-dihydroxy-2-(3-methy-lbut-2-enyl)-3-methoxyxanthone(3), cratoxyxanthone(4), 2,6-dimethoxy-para-benzoquinone(5), methyl orselinate(6), ficusol(7), and 4-(4-carboxy-2-methoxyphenoxy)-3,5-dimethoxybenzoic acid(8). Compound 1 was a new xanthone, and compound 4 was a xanthone dimer, compound 5 was a naphthoquinone. All compounds were isolated from this plant for the first time except compounds 2 and 3. Cytotoxic bioassay suggested that compounds 1, 2 and 4 possessed moderate cytotoxicity, suppressing HeLa cell line with IC_(50) va-lues of 24.3, 35.5 and 17.1 μmol·L~(-1), respectively. Compound 4 also could suppress K562 cells with an IC_(50) value of 39.8 μmol·L~(-1).
Humans
;
Garcinia mangostana/chemistry*
;
HeLa Cells
;
Antineoplastic Agents
;
Magnetic Resonance Spectroscopy
;
Xanthones/pharmacology*
;
Garcinia/chemistry*
;
Plant Extracts/chemistry*
;
Molecular Structure
7.Preparation of an ophthalmic formulation of TPGS-modified insulin-loaded liposomes and its <i>in vitroi> corneal permeation and pharmacokinetics in rabbit eyes.
Journal of Southern Medical University 2023;43(5):832-838
OBJECTIVE:
To prepare vitamin E polyethylene glycol 1000 succinate (TPGS)-modified insulin-loaded liposomes (T-LPs/INS) and evaluate its safety, corneal permeability, ocular surface retention and pharmacokinetics in rabbit eyes.
METHODS:
The safety of the preparation was investigated in human corneal endothelial cells (HCECs) using CCK8 assay and live/dead cell staining. In the ocular surface retention study, 6 rabbits were randomized into 2 equal groups for application of fluorescein sodium dilution or T-LPs/INS labeled with fluorescein in both eyes, which were photographed under cobalt blue light at different time points. In the cornea penetration test, another 6 rabbits divided into 2 groups for application of Nile red diluent or T-LPs/INS labeled with Nile red in both eyes, after which the corneas were harvested for microscopic observation. In the pharmacokinetic study, 2 groups of rabbits (<i>ni>=24) were treated with eye drops of T-LPs/INS or insulin, and the aqueous humor and cornea were collected at different time points for measurement of insulin concentrations using enzyme linked immunosorbent assay. DAS2 software was used to analyze the pharmacokinetic parameters.
RESULTS:
The prepared T-LPs/INS showed good safety in cultured HCECs. Corneal permeability assay and fluorescence tracer ocular surface retention assay demonstrated a significantly higher corneal permeability of T-LPs/INS with a prolonged drug residence in the cornea. In the pharmacokinetic study, insulin concentrations in the cornea at 6, 15, 45, 60, and 120 min (<i>Pi> < 0.01) and in the aqueous humor at 15, 45, 60, and 120 min after dosing were significantly higher in T-LPs/INS group. The changes in insulin concentrations in the cornea and aqueous humor were consistent with a two-compartment model in T-LPs/INS group and with the one-compartment model in the insulin group.
CONCLUSION
The prepared T-LPs/INS shows an improved corneal permeability, ocular surface retention capacity and eye tissue concentration of insulin in rabbits.
Humans
;
Animals
;
Rabbits
;
Insulin
;
Liposomes
;
Endothelial Cells
;
Lipopolysaccharides
;
Vitamin E
;
Cornea
;
Fluorescein
8.Regulatory Mechanism of Mangiferin Combined with Bortezomib on Malignant Biological Behavior of Burkitt Lymphoma and Its Effect on Expression of CXC Chemokine Receptors.
Zhi-Min YAN ; Yan-Quan LIU ; Qing-Lin XU ; Jie LIN ; Xin LIU ; Qiu-Ping ZHU ; Xin-Ji CHEN ; Ting-Bo LIU ; Xiao-Lan LIAN
Journal of Experimental Hematology 2023;31(5):1394-1402
OBJECTIVE:
To analyze the effects of mangiferin combined with bortezomib on the proliferation, invasion, apoptosis and autophagy of human Burkitt lymphoma Raji cells, as well as the expression of CXC chemokine receptors (CXCRs) family, and explore the molecular mechanism between them to provide scientific basis for basic research and clinical work of Burkitt lymphoma.
METHODS:
Raji cells were intervened with different concentrations of mangiferin and bortezomib alone or in combination, then cell proliferation was detected by CCK-8 assay, cell invasion ability was detected by Transwell chamber method, cell apoptosis was detected by Annexin V/PI double-staining flow cytometry, apoptosis, autophagy and Akt/mTOR pathway protein expression were detected by Western blot, and the expression changes of CXCR family was detected by real-time quantitative PCR (RT-qPCR).
RESULTS:
Different concentrations of mangiferin intervened Raji cells for different time could inhibit cell viability in a concentration- and time-dependent manner (<i>ri> =-0.682, <i>ri> =-0.836). When Raji cells were intervened by combination of mangiferin and bortezomib, compared with single drug group, the proliferation and invasion abilities were significantly decreased, while the apoptosis level was significantly increased (<i>Pi> <0.01). Mangiferin combined with bortezomib could significantly up-regulate the expression of pro-apoptotic protein Bax and down-regulate the expression of anti-apoptotic protein Bcl-2 after intervention in Raji cells. Caspase-3 was also hydrolyzed and activated, and then induced the apoptosis of Raji cells. Mangiferin combined with bortezomib could up-regulate the expression of LC3Ⅱ protein in Raji cells, and the ratio of LC3Ⅱ/LC3Ⅰ in cells was significantly up-regulated compared with single drug or control group (<i>Pi> <0.01). Mangiferin combined with bortezomib could significantly inhibit the phosphorylation levels of Akt and mTOR, inhibit the proliferation and invasion of Raji cells by inhibiting Akt/mTOR pathway, and induce cell autophagy and apoptosis. Mangiferin and bortezomib could down-regulate the expressions of <i>CXCR4i> and <i>CXCR7i> mRNA after single-agent intervention in Raji cells, and the down-regulations of <i>CXCR4i> and <i>CXCR7i> mRNA expression were more significant when the two drugs were combined (<i>Pi> <0.01). Mangiferin alone or combined with bortezomib had no significant effect on <i>CXCR5i> mRNA expression in Raji cells (<i>Pi> >0.05), while the combination of the two drugs could down-regulate the expression of <i>CXCR3i> (<i>Pi> <0.05).
CONCLUSION
Mangiferin combined with bortezomib can synergistically inhibit the proliferation and invasion of Raji cells, and induce autophagy and apoptosis. The mechanism may be related to the inhibition of Akt/mTOR signaling pathway, down-regulation of anti-apoptotic protein Bcl-2 and up-regulation of pro-apoptotic protein Bax, and the inhibition of the expression of CXCR family.
Humans
;
Antineoplastic Agents/therapeutic use*
;
Apoptosis/drug effects*
;
Apoptosis Regulatory Proteins/immunology*
;
Autophagy/immunology*
;
bcl-2-Associated X Protein/immunology*
;
Bortezomib/therapeutic use*
;
Burkitt Lymphoma/immunology*
;
Cell Line, Tumor
;
Cell Proliferation/drug effects*
;
Drug Therapy, Combination
;
Proto-Oncogene Proteins c-akt
;
Proto-Oncogene Proteins c-bcl-2
;
Receptors, CXCR/immunology*
;
RNA, Messenger
;
TOR Serine-Threonine Kinases
;
Xanthones/therapeutic use*
9.Application of Rapid HE Staining in Cytological Rapid On-site Evaluation of Peripheral Lung Cancer Needle Biopsy.
Jian HE ; Guilan XIA ; Shiping WANG ; Kun CHEN
Chinese Journal of Lung Cancer 2023;26(8):572-578
BACKGROUND:
Rapid on-site evaluation (ROSE) is a technique used for simultaneous evaluation of biopsy specimens through rapid cytology staining. Diff-Quik (DQ) staining is the most commonly employed method for cytological rapid on-site evaluation (C-ROSE). However, the utilization of DQ staining for on-site cytological interpretation remains uncommon among pathologists in China, posing challenges to the implementation of C-ROSE. This study aims to assess the application of rapid hematoxylin-eosin (HE) staining and DQ staining for C-ROSE during percutaneous needle biopsy of peripheral lung cancer and evaluate the value of rapid HE staining in C-ROSE.
METHODS:
Computed tomography (CT)-guided lung biopsies were conducted on 300 patients diagnosed with peripheral lung cancer. The patients were randomly assigned to two groups for C-ROSE using either rapid HE staining or DQ staining, and subsequently the two methods were compared and evaluated.
RESULTS:
The concordance rate between C-ROSE and histopathological diagnosis was 96.7%. The median staining time for rapid HE staining was 160 s, while that for DQ staining was 120 s, representing a significant difference between the two groups (P<0.001). However, there were no significant differences observed in terms of total biopsy time, concordance rate with histopathology, cytology specimen peeling rate, and incidence of serious adverse reactions between the two groups (P>0.05).
CONCLUSIONS
Both staining methods comply with C-ROSE criteria in the biopsy setting of peripheral lung cancer. Rapid HE staining is more aligned with domestic clinical requirements and holds potential for further promotion and adoption in C-ROSE.
Humans
;
Lung Neoplasms/pathology*
;
Eosine Yellowish-(YS)
;
Rapid On-site Evaluation
;
Biopsy, Needle/methods*
;
Staining and Labeling
10.Effect of P311 microspheres-loaded thermosensitive chitosan hydrogel on the wound healing of full-thickness skin defects in rats.
Qing Rong ZHANG ; Chang You CHEN ; Na XU ; Da Lun LYU ; Jie Zhi JIA ; Wen Hong LI ; Gao Xing LUO ; Yun Long YU ; Yi ZHANG
Chinese Journal of Burns 2022;38(10):914-922
Objective: To explore the effect of P311 microspheres-loaded thermosensitive chitosan hydrogel on the wound healing of full-thickness skin defects in rats. Methods: The method of experimental study was adopted. The polyvinyl alcohol/sodium alginate microspheres (simple microspheres), P311 microspheres, and bovine serum albumin labeled with fluorescein isothiocyanate (FITC-BSA) microspheres were prepared by water-in-oil emulsification, and then their morphology was observed under a light microscope/inverted fluorescence microscope. Chitosan solution was prepared, chitosan solution and β-glycerol phosphate disodium hydrate were mixed to prepare simple thermosensitive hydrogels, and thermosensitive hydrogels loaded with simple microspheres or P311 microspheres were prepared by adding corresponding substances in simple thermosensitive hydrogels. The morphological changes of the prepared four liquids in the state of tilt was observed at 37 ℃. After being freeze-dried, the micromorphology of the prepared four liquids was observed under a scanning electron microscope. Eighteen 3-4-week-old male Sprague-Dawley rats were divided into normal group without any treatment, dressing group, chitosan group, hydrogel alone group, simple microspheres-loaded hydrogel group, and P311 microspheres-loaded hydrogel group, which were inflicted with one full-thickness skin defect wound on both sides of the back spine and were dealt correspondingly, with 3 rats in each group. Rats with full-thickness skin defects in the five groups were collected, the wound healing was observed on post injury day (PID) 0 (immediately), 5, 10, and 15, and the wound healing rates on PID 5, 10, and 15 were calculated. The wound and wound margin tissue of rats with full-thickness skin defects in the five groups on PID 15 and normal skin tissue in the same site of rats in normal group were collected, hematoxylin and eosin staining was conducted to observe the histological changes, immunohistochemical staining was performed to observe the expressions of CD31 and vascular endothelial growth factor (VEGF), and Western blotting was conducted to detect the protein expressions of CD31 and VEGF. The number of samples was all three. Data were statistically analyzed with one-way analysis of variance, analysis of variance for repeated measurement, and Bonferroni correction. Results: Simple microspheres were spherical, with loose and porous surface. The surfaces of P311 microspheres and FITC-BSA microspheres were smooth without pores, and the FITC-BSA microspheres emitted uniform green fluorescence. The diameters of the three microspheres were basically consistent, being 33.1 to 37.7 μm. Compared with chitosan solution and simple thermosensitive hydrogel, the structures of the two microspheres-loaded hydrogels were more stable in the state of tilt at 37 ℃. The two microspheres-loaded hydrogels had denser network structures than those of chitosan solution and simple thermosensitive hydrogel, and in the cross section of which microspheres with a diameter of about 30 μm could be seen. Within PID 15, the wounds of rats in the five groups were healed to different degrees, and the wound healing of rats in P311 microspheres-loaded hydrogel group was the best. On PID 5, 10, and 15, the wound healing rates of rats in dressing group and chitosan group were (26.6±2.4)%, (38.5±3.1)%, (50.9±1.5)%, (47.6±2.0)%, (58.5±3.6)%, and (66.7±4.1)%, respectively, which were significantly lower than (59.3±4.8)%, (87.6±3.2)%, (97.2±1.0)% in P311 microspheres-loaded hydrogel group (<i>Pi><0.05 or <i>Pi><0.01). The wound healing rates of rats in hydrogel alone group on PID 10 and 15, and in simple microspheres-loaded hydrogel group on PID 15 were (76.0±3.3)%, (84.5±3.6)%, and (88.0±2.6)%, respectively, which were significantly lower than those in P311 microspheres-loaded hydrogel group (<i>Pi><0.05). The epidermis, hair follicles, and sebaceous glands could be seen in the normal skin of rats in normal group, without positive expressions of CD31 or VEGF. The wounds of rats in P311 microspheres-loaded hydrogel group on PID 15 were almost completely epithelialized, with more blood vessels, hair follicles, sebaceous glands, and positive expressions of CD31 and VEGF in the wounds than those of rats with full-thickness skin defects in the other four groups, and more protein expressions of CD31 and VEGF than those of rats in the other five groups. Conclusions: The P311 microspheres-loaded thermosensitive chitosan hydrogel can release the encapsulated drug slowly, prolong the drug action time, and promote wound healing in rats with full-thickness skin defects by promoting wound angiogenesis and re-epithelialization.
Rats
;
Male
;
Animals
;
Hydrogels
;
Vascular Endothelial Growth Factor A
;
Chitosan/pharmacology*
;
Serum Albumin, Bovine/pharmacology*
;
Microspheres
;
Polyvinyl Alcohol/pharmacology*
;
Hematoxylin/pharmacology*
;
Eosine Yellowish-(YS)/pharmacology*
;
Rats, Sprague-Dawley
;
Wound Healing
;
Skin/injuries*
;
Skin Abnormalities
;
Soft Tissue Injuries
;
Water/pharmacology*
;
Alginates/pharmacology*


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