Elsevier

Bone

Volume 35, Issue 4, October 2004, Pages 892-898
Bone

Transplantation of marrow-derived mesenchymal stem cells and platelet-rich plasma during distraction osteogenesis—a preliminary result of three cases

https://doi.org/10.1016/j.bone.2004.06.013Get rights and content

Abstract

Clinical results of distraction osteogenesis with transplantation of marrow-derived mesenchymal stem cells (MSCs) and platelet-rich plasma (PRP) were reviewed in three femora and two tibiae of the two patients with achondroplasia and one patient with congenital pseudarthrosis of the tibia. MSCs derived from the iliac crest were cultured with osteogenic supplements and differentiated into osteoblast-like cells. PRP, which is known to contain several growth factors and coagulate immediately by a minute introduction of thrombin and calcium, was prepared just before transplantation. Culture-expanded osteoblast-like cells and autologous PRP were injected into the distracted callus with the thrombin–calcium mixture so that the PRP gel might develop within the injected site. Transplantation of MSCs and PRP was done at the lengthening and consolidation period in each patient. The target lengths were obtained in every leg without major complications and the average healing index was 23.0 days/cm (18.8–26.9 days/cm). Although these results are still preliminary, transplantation of osteoblast-like cells and PRP, which seemed to be a safe and minimally invasive cell therapy, could shorten the treatment period by acceleration of bone regeneration during distraction osteogenesis.

Introduction

Distraction osteogenesis has been successfully used for limb lengthening in patients with limb length discrepancy, severe short stature due to skeletal dysplasias, and large bone defect that develop as a result of etiological reasons such as tumor, infection, and trauma. Although this treatment method is biological and has many advantages for these pathological conditions, the periods of external fixation and bone maturation is long which results in higher rates of complications such as pin track infection, joint contractures, pin loosening, delayed consolidations and fractures [1]. Decreasing the treatment period by accelerating bone formation of the distracted callus could reduce these complications.

Bone marrow contains a population of multipotent mesenchymal stem cells (MSCs) that generate the progenitors for osteogenic, chondrogenic, adipocytic, and myogenic cells [2]. MSCs can be directed towards the osteogenic lineage in vitro if cultured in the presence of dexamethasone, β-glycerophosphate and ascorbic acid. Furthermore, MSCs are appropriate for donor tissue and may be used in clinical applications because they are easily isolated from a small aspirate of bone marrow and readily generate various cell types. Using the rat limb lengthening model, we have previously demonstrated that transplantation of marrow derived osteoblast-like cells with collagen gel into the distracted callus promoted new bone formation and shortened the consolidation period [3].

Biodegradable carriers or scaffolds are needed for MSCs transplantation and tissue growth. Heterologous or allogeneic materials are not appropriate for clinical feasibility because it carries potential risks of infection, immune responses and pathogen transmission. Platelet-rich plasma (PRP), which is developed from autologous blood with a cell separator, is rich in platelets and contains several growth factors including platelet-derived growth factor (PDGF), insulin-like growth factor (IGF-1), and transforming growth factors (TGF-β1 and β2) [4], [5]. These growth factors enhance and accelerate the normal bone regeneration pathways [6]. Autologous PRP could be a suitable carrier for MSCs transplantation because it coagulates immediately by a minute introduction of calcium and thrombin.

In this paper, we introduce a new cell therapy during distraction osteogenesis using culture expanded MSCs and autologous PRP. Preliminary results of the limb lengthening procedures with MSCs and PRP were reviewed.

Section snippets

Patients

After informed consent was obtained from all individuals before surgery, 10 limb lengthening procedures with transplantation of MSCs and PRP were performed in seven patients at the Nagoya University between July 2002 and August 2003. Four femoral and two tibial lengthenings were performed in three achondroplasia patients bilaterally, and three femoral and one tibial lengthenings were done in four patients because of a limb length discrepancy, which was secondary to trauma (two limbs),

Characterization of MSCs

After several days in culture, a low number of fibroblast-like appearance MSCs was obtained, although the initial adherence of MSCs was diverse in each case. On approximately day 10, MSCs grew to reach a semiconfluent monolayer in primary culture with the average of 1.2 × 106 cells. The suspended cells reached confluence in 3–5 days and the third-passaged (P3) cells with the average of 2.3 × 107 (1.4 × 107–5.0 × 107) were used for transplantation. Under the culture conditions with osteogenic

Discussion

The incidence and severity of complications during limb lengthening tends to increase in relation to the period of external fixation. Many attempts using electronic stimulation, hyperbaric oxygen exposure, low intensity pulsed ultrasound stimulation, or systemic administration of recombinant growth hormone, have been made to promote bone formation during consolidation period [7], [8], [9], [10]. Enhanced new bone regenerates have been demonstrated in rabbit models of distraction osteogenesis by

Acknowledgment

This work was supported in part by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan (contact grant number 14370461 and 15591573).

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