Potential of Continuous Local Antibiotic Perfusion Therapy for Fracture-Related Infections: A Case Series

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Additional file 1 of DAIR in treating chronic PJI after total knee arthroplasty using continuous local antibiotic perfusion therapy: a case series study

Supplementary Material 1: Fig. A Dual-lumen tubes (Salam samp tube®: Nihon Covidien Co.

Nano-fiber plugs induce osteogenesis of human MSCs.

<p>Healthy donor-derived MSCs were seeded onto plastic plates (A-C) or injected into the nano-fiber plugs (D-I), and then cultured in MSCGM for 1 day (D), 7 days (E, G), 14 days (F, H) and 28 days (A-C, I). Then, the expression levels of RUNX2 (A, D, G), osteocalcin (B, E, H), and DMP-1 (C, F, I) were evaluated immunohistochemically. Representative results of three experiments are shown. (J) Healthy donor-derived MSCs were cultured in MSCGM onto plastic plates (two-dimensional) or nano-fiber plugs (three-dimensional) for 28 days. After RNA extraction, <i>MEPE</i> expression in the 4 groups of healthy donor-derived MSCswas evaluated by real-time PCR. *p<0.05 vs. two-dimensional culture by paired <i>t</i>-test. Scale bars, 50 μm.</p

The hierarchy of gene expression in MSCs during differentiaion into osteoblasts, chondrocytes, and osteocytes.

<p>MSCs differentiate into osteoblasts or chondrocytes under the regulation of their master regulators, RUNX2 or SOX9, respectively. A part of chondrocytes also differentiate into osteoblasts.</p

Effects of lactic acid on cell functions of human MSCs.

Healthy donor-derived MSCs seeded onto plastic plates were cultured in MSCGM with lactic acid. (A) Proliferative capacity assessed by measurement of 3H thymidine uptake after 1 day (n = 3). (B) Propidium iodide-positive apoptotic cells and WST-8-positive viable cells on Day 7 (n = 5). (C) ALP activity on Day 7 (n = 3), and (D) production of sGAG on Day 28 (n = 3). Data are mean±SD. *p<0.05, **p<0.01 vs. 0 mM, by ANOVA with post-hoc Dunnett’s test. n.s.: not significant.</p

Nano-fiber plugs induce chondrogenesis of human MSCs.

(A) Healthy donor-derived MSCs were injected into the center of nano-fiber plugs and cultured in MSCGM for the specified period. The samples were assessed immunohistochemically for the expression of type-II collagen on Days 7 and 28. Representative results of three experiments are shown. Scale bars, 50 μm. (B and C) Human MSCs were cultured in MSCGM onto plastic plates (two-dimensional) or nano-fiber plugs (three-dimensional) for the specified period. After RNA extraction, the expression levels of SOX9 at day 7 (B) and COL10A1 at day 28 (C) in the 4 groups of healthy donor-derived MSCswere assessed by real-time PCR. *pt-test.</p

MSCs cultured on nano-fiber plugs produce minerals and proteoglycan upon differentiation stimulation.

<p>Healthy MSCs were seeded onto plastic plates (A, D) or injected into the center of the nano-fiber plugs (B, C, E, F), and then cultured in OIM (A-C) or CM (D-F) for 28 days. The samples were then fixed with formalin, and then stained with von Kossa stain (A, B, E) or Safranin O (C, D, F). Representative results of three experiments are shown. Scale bars, 50 μm.</p

Nano-fiber plugs induce MSCs to produce minerals and proteoglycan.

<p>Healthy donor-derived MSCs (A-F) or healthy donor-derived skin fibroblasts (G, H) were seeded onto plastic plates (A, B) or injected into the center of the nano-fiber plugs (C-H), and then cultured in mesenchymal stem cell growth medium for 14 days (C, E), 28 days (A, B, F-H), or 56 days (D). The samples were fixed with formalin, embedded in paraffin, and stained with von Kossa stain (A, C, D, G) or Safranin O (B, E, F, H). Representative results of three experiments are shown. Scale bars, 50 μm.</p

Nano-fiber plugs induce the differentiation of MSCs derived from patients with RA or OA.

<p>MSCs derived from patients with RA (n = 3) or OA (n = 3) were cultured in MSCGM onto plastic plates (two-dimensional) or nano-fiber plugs (three-dimensional) for 7 (A) or 28 days (B). After RNA extraction, real-time PCR was performed to evaluate the gene expression level. Each line represents one patient. MSCs from healthy donors (n = 6), RA (n = 3) or OA (n = 3) patients were cultured in NFs for 7 days (C) or 28 days (D), and then analyzed as for their gene expression by real-time PCR. n.s.: not significant by ANOVA.</p

Nano-fiber plugs induce morphological changes in MSCs.

<p>Healthy donor-derived MSCs were seeded onto nano-fiber plugs, and then cultured in MSCGM. Scanning electron micrographs taken on Days 7 (A) and 28 (B) are shown. All samples were tested three times, and representative results are shown. Scale bars, 50 μm.</p