Autologous bone marrow stem cell intralesional transplantation repairing bisphosphonate related osteonecrosis of the jaw
© Cella et al; licensee BioMed Central Ltd. 2011
Received: 24 May 2011
Accepted: 17 August 2011
Published: 17 August 2011
Bisphosphonate - related osteonecrosis of the JAW (BRONJ) is a well known side effect of bisphosphonate therapies in oncologic and non oncologic patients. Since to date no definitive consensus has been reached on the treatment of BRONJ, novel strategies for the prevention, risk reduction and treatment need to be developed. We report a 75 year old woman with stage 3 BRONJ secondary to alendronate and pamidronate treatment of osteoporosis. The patient was unresponsive to recommended treatment of the disease, and her BRONJ was worsening. Since bone marrow stem cells are know as being multipotent and exhibit the potential for differentiation into different cells/tissue lineages, including cartilage, bone and other tissue, we performed autologous bone marrow stem cell transplantation into the BRONJ lesion of the patient.
Under local anesthesia a volume of 75 ml of bone marrow were harvested from the posterior superior iliac crest by aspiration into heparinized siringes. The cell suspension was concentrated, using Ficoll - Hypaque® centrifugation procedures, in a final volume of 6 ml. Before the injection of stem cells into the osteonecrosis, the patient underwent surgical toilet, local anesthesia was done and spongostan was applied as a carrier of stem cells suspension in the bone cavity, then 4 ml of stem cells suspension and 1 ml of patient's activated platelet-rich plasma were injected in the lesion of BRONJ.
A week later the residual spongostan was removed and two weeks later resolution of symptoms was obtained. Then the lesion improved with progressive superficialization of the mucosal layer and CT scan, performed 15 months later, shows improvement also of bone via concentric ossification: so complete healing of BRONJ (stage 0) was obtained in our patient, and 30 months later the patient is well and without signs of BRONJ.
To our knowledge this is the first case of BRONJ successfully treated with autologous stem cells transplantation with a complete response.
KeywordsOsteonecrosis of the Jaw bisphoshonate stem cell transplantation organ repair
Bisphosphonates are widely used in the management of bone diseases including osteoporosis, Paget's disease, hypercalcemia related to malignancy and in the prevention of skeletal complication from bone metastasis. Bisphosphonates are incorporated into skeleton and suppress bone resorption, without being degraded [1, 2]. Bisphosphonates have shown direct anti-tumor effects, possibly related to growth factors release reduction and inhibition of cell adhesion molecules [2, 3]. Bisphosphonates - related osteonecrosis of the Jaw (BRONJ) has been characterized as a main side effect of bisphosphonate therapy [4–6]. The most frequent clinical sign of BRONJ is bone exposure, associated with pain, swelling and purulent secretion that does not heal over a period of 6-8 weeks [7, 8]. To date no definitive consensus has been reached on the treatment of BRONJ and several studies reported relatively conflicting results following surgery, antibiotics, laser or hyperbaric oxygen administration [9–16]. For this reason, new strategies for the prevention, risk reduction and treatment for BRONJ need to be developed [16, 14, 17–20].
Bone marrow harvested stem cells and progenitor cells (BMSC) may be capable of solid-organ repair , and it has been demonstrated that adult human mesenchymal stem cells (MSC) from bone marrow can represent a promising source for skeletal regeneration .
Based on these data, we report here a patient with BRONJ, unresponsive to the recommended procedures, that showed clinical and radiographic improvement after autologous bone marrow stem cells intralesional transplantation.
Current or previous treatment with a bisphosphonate;
Exposed bone in the maxillofacial region that has persisted fore more 8 weeks;
No history of radiation therapy to the jaws.
The concept that bone marrow stem cells upon transplantation into adult recipients transdifferentiate and contribute to the rigeneration of a variety of non -hematopoietic lineages in multiple organs, has provoked great interest for its potential clinical applications [24, 25] as recently reported also by our group . So we offered the opportunity to our patient of injection of autologous bone marrow stem cells into the osteonecrosis site lesion.
In september 2008 the patient (who is the mother of one of us) was informed about the procedure and gave written informed consent.
Under local anesthesia an average of 75 ml of bone marrow were harvested from the posterior superior iliac crest by aspiration into heparinized syringes as previously reported by our group . Progenitor cells were isolated and enriched using Ficoll - Hypaque® centrifugation procedures. This procedure allowed the depletion of mature myeloid and erythroid cell from the harvest. The cell suspension consisted of an heterogeneous cell population including hematopoietic, mesenchymal, endothelial and other progenitor cells as well as mononuclear cells. The cells were suspended into an opportune quantity of PBS-EDTA buffer containing 5% of human albumin.
Multiparameter flow cytometric analysis of the injected BMC
harvest (ml 75)
final (ml 6)
injected (ml 4)
nuclear cells/ul (total E6)
WBC (CD45+) (E6) *
CD 34+ cells/ul
CD 34+ cells (E6)
CD34+/CD117+ cells (E6)
CD34+/CD133+ cells (E6)
CD45-/CD31+ cells (E6)
CD133+/CD117+ cells (E6)
CD133+ cells (E6)
CD117+ cells (E6)
CD45-/CD105+/CD71- cells (E6)
Ficoll mediated myeloid depletion % *
Before the injection of stem cells into the osteonecrosis the patient underwent a surgical toilet in local anesthesia of the bone lesion.
The bone cavity was fullfilled with fibrine sponge (Spongostan®) as a carrier, then 4 ml of stem cells suspension and 1 ml of patient's activated platelet-rich plasma were injected in the lesion of BRONJ.
Since the first description by Marx, 2003 and Wang et al 2003 , cases with BRONJ are being increasingly reported, first of all, in oncologic patients in line with the increased use of bisphosphonates (mainly zolendronate and pamidronate) as the main pathogenetic factor of BRONJ.
A review of the literature through march 2006 performed by our group  identified more than 250 reported case on BRONJ, and more recently over 6.000 cases have been reported to the US Food and Drug Administration . The treatment goals for patients with an established diagnosis of BRONJ are, as recently reported [16–20], to eliminate pain, to control infection of the soft and hard tissue and to minimize the occurrence or the progression of bone necrosis.
However the response to treatments of the patients with BRONJ is less predictable than the established surgical treatment modalities for osteomyelitis or osteoradionecrosis, and new treatment procedures need to be developed [16–20].
From the hystopathological point of view the BRONJ is characterized by an avascular necrosis. In osteonecrosis is found a lack of osteogenic precursor cells that derive from mesenchymal stem cells (MSCs), but also a lack in vascular support that derives from endothelial progenitor cells (EPCs).
MSCs are known as being multipotent and exhibit the potential for differentiation into different cell/tissue lineages, including cartilage, bone, adipose tissue, tendon and ligament . These pluripotent mesenchymal progenitor cell are denoted as stromal or mesenchymal stem cells.
In vivo osteogenesis occurs only if the density of implanted cells at the treated site is sufficiently high. To achieve this goal, either large amounts of concentrated bone marrow stem cells or bone marrow stem cells in combination with growth factors can be used [31, 32].
This situation has been reproduced by in vitro studies which confirmed that composite implantation of mesenchymal stem cells with endothelial progenitor cells enhances tissue-engineered bone formation .
The homing mechanisms of MSCs are poorly understood; it is known that, based on chemokine/chemokine-receptor interactions and adhesion molecules, MSCs are potentially capable on finding the site of injury and when, given intravenously, of restoring damaged tissue on site due to their plasticity and/or paracrine properties . However, it must be emphasized that a direct approach, bringing direct into the osteonecrotic site a significant amount of bone marrow enriched in mononuclear cells, could allow a better osteogenesis of the damaged bone based on evidence data of the presence in this cell-fraction of osteoid and angiogenic precursors [34–36].
Bone marrow contains three main cell lines: hematopoietic cells, mesenchymal and proendotelial cells [34, 35]. Recently we reported in a randomized controlled trial the effects of intracoronary transfer of autologous bone marrow stem cells in patients with acute anterior myocardial infarction and we demonstrated that this procedure improves cardiac, autonomic, and functional indexes in this setting of treated patients . These positive effects may be mediated by a direct transdifferentiation of transplanted stem cells to cardiomyocytes , but also indirectly by parackrine secretion of cytokines and growth factors with resulting stimulation of survivors cardiac stem cells and/or angiogenesis, improving microvascular function . Recently, a stabilizing effect of the injection cells via changes in the connettive tissue has been hypothesized .
Stem cells are easily obtained from the bone marrow with a minimally invasive approach and can be easily transplanted into the osteonecrotic lesion as demonstrated in our patient. This simple, cheap procedure allowed a clinical improvement of symptoms, and induced novel ossification as demonstrated by CT scan 15 months after the treatment, and it must be emphasized that the patient is in complete remission from a stage 3 BRONJ after 30 months. In addition, our patient showed a particularly rich bone marrow, not only in red cells precursors (as we expected since the patient was treated with EPO), but also in total stem cells subset (table 1). Recently, Kikuiri et al,  infused mesenchymal stem cells in BRONJ-like mice. They demonstrated that systemic infusion with MSCs prevents and cures BRONJ-like disease possibly via introduction of peripheral tolerance, shown as an inhibition of T-helper-producing interlukin 17 cells (th17)and increase in T regulatory cells (Tregs). Handschel and Meyer  suggest that stem cells might be a promising treatment option for BRONJ and our case demonstrates their hypothesis is right. In our case bone marrow stem cells were directly infused in the bone lesion of BRONJ with a complete remission.
We are aware that a case report can be of limited interest, however it could suggest that this technique may be studied in patients with BRONJ unrensponsive to standard treatment and can be tried before major demolitive surgery procedures for the reconstruction of defect of the ONJ by bisphosphonates.
Written informed consent was obtained from the patient for publication of this case report and accompanying images. A copy of the written consent is available for review by the Editor-in-Chief of this journal
Authors acknowledge Fondazione di Piacenza e Vigevano (Italy) for the excellent support and assistence
- McClug MR: Bisphosphonates. Endocrinol Metab Clin North Am. 2003, 32: 253 71Google Scholar
- Santini D, Vespasiani Gentilucci U, Vincenzi B, Picardi A, Vasaturo F: The antineoplastic role of bisphosphonates: from basic research to clinical evidence. Ann Oncol. 2003, 14: 1468-76. 10.1093/annonc/mdg401.View ArticlePubMedGoogle Scholar
- Boissier S, Magnetto S, Frappart L, Cuzin B, Ebetino FH, Delmas PD: Bisphosphonates inhibit prostate and breast carcinoma cell adhesion to unmineralized and mineralized bone extracellular matrices. Cancer Res. 1997, 57: 3890-3894.PubMedGoogle Scholar
- Wysoski DK: Reports of esophageal cancer with oral bisphosphonates use. N Engl J Med. 2009, 360: 89-90. 10.1056/NEJMc0808738.View ArticleGoogle Scholar
- Ibrahim T, Barbanti F, Giorgio Marrano G, Mercatali L, Ronconi S, Vicini C: Osteonecrosis of the Jaw in patients with bone matastases treated with bisphosphonates: a retrospective study. Oncologist. 2008, 13: 330-336. 10.1634/theoncologist.2007-0159.View ArticlePubMedGoogle Scholar
- Bagan JV, Jimenez Y, Murillo J, Hernandez S, Poveda R, Sanchis JM: Jaw osteonecrosis associated with bisphosphonates: multiple exposed areas and its relationship to teeth extractions. Study of 20 cases. Oral oncol. 2006, 42: 327-329. 10.1016/j.oraloncology.2005.08.001.View ArticlePubMedGoogle Scholar
- Hewitt C, Farah C: Bisphosphonates-related osteonecrosis of the jaws: a comprehensive review. J Oral Pathol Med. 2007, 36: 319-328. 10.1111/j.1600-0714.2007.00540.x.View ArticlePubMedGoogle Scholar
- American Dental Association Council on Scientific Affairs: Dental Management of patients receiving oral bisphosphonate Therapy: Expert panel recommendations. J Am Assoc. 2006, 137: 1144-1150.Google Scholar
- Cavanna L, Bertè R, Arcari A, Mordenti P, Pagani R, Vallisa D: Osteonecrosis of the Jaw. A newly emerging sete-specific osseous pathology in opatients with cancer treated with bisphosphonates. Report of 5 cases and review of the literature. Eur J Internal Med. 2007, 18: 417-422. 10.1016/j.ejim.2006.10.008.View ArticleGoogle Scholar
- Rizzoli R, Burlet N, Cahall D, Delmas PD, Eriksen EF, Felsenberg D: Osteonecrosis of the jaw and bisphosphonate treatment for osteoporosis. Bone. 2008, 42: 841-847. 10.1016/j.bone.2008.01.003.View ArticlePubMedGoogle Scholar
- Marx RE, Sawatari Y, Fortin M, Broumand V: Bisphosphonate-induced exposed bone (osteonecrosis/osteopetrosis) of the jaws: risk factors, recognition, prevention and treatment. J Oral Maxillofac Surg. 2005, 63: 1567-1575. 10.1016/j.joms.2005.07.010.View ArticlePubMedGoogle Scholar
- Yarom N, Yahalom R, Shoshani Y, Hamed W, Regev E, Elad S: Osteonecrosis of the jaw induced by orally administered bisphosphonates: incidence, clinical features, predisposing factors and teatment outcome. Osteoporos Int. 2007, 18: 1363-1370. 10.1007/s00198-007-0384-2.View ArticlePubMedGoogle Scholar
- Elad S, Yarom N, Hamed W, Ayalon S, Yahalom R, Regev E: Osteomyelitis and necrosis of the jaw in patients treated with bisphosphonates A comparative study. Clin Lab Haematol. 2006, 28: 393-398. 10.1111/j.1365-2257.2006.00841.x.View ArticlePubMedGoogle Scholar
- Ruggiero SL, Fantasia J, Carlson E: Bisphosphonates-related osteonecrosis of the jaw: background and guidlines for diagnosis, staging and management. Oral Surg. Oral Med Oral Path Oral Radio Endod. 2006, 102: 433-41. 10.1016/j.tripleo.2006.06.004.View ArticleGoogle Scholar
- Vescovi P, Merigo E, Meleti M, Fornaini C, Bonanini M, Rocca EP, De Moor RJ, Nammour S: Surgical treatment of maxillary osteonecrosis due to bisphosphonates using an Er: YAG (2940 nm) laser. Discussion of 17 clinical cases. Rev Belge Med Dent. 2009, 64 (2): 87-95.Google Scholar
- Rugani P, Stephan A, Truschnegg A, Obermayer-Pietsch B, Jakse N: Bisphosphonate - associated osteonecrosis of the jaws: surgical treatment with ErCrYSGG-laser. Case report. OOOOE. 2010, 110: e1-e6.PubMedGoogle Scholar
- Ruggiero SL, Dodson TB, Assale LA, Landesberg R, Marx RE, Mehrotra B: American Association of Oral and Maxillofacial Surgeons Position Paper on Bisphosphonate-Relate Osteonecrosis of the Jaws-2009 Update. J Oral Maxillofac Surg. 2009, 67 (Suppl): 2-12.PubMedGoogle Scholar
- Madrid C, Bouferrache K, Abarca M, Jaques B, Broome M: Bisphosphonate-related osteonecrosis of the jaws: how to manage cancer patients. Oral Oncology. 2010, 46: 468-470. 10.1016/j.oraloncology.2010.03.016.View ArticlePubMedGoogle Scholar
- Lee JJ, Cheng SJ, Jeng JH, Ching CP, Lau HP, Kok SH: Successful treatment of advanced bisphosphonate-related osteonecrosis of the mandible with adjunctive teriparatide therapy. Wiley InterScience. 2010, [http://www.interscience.wiley.com]Google Scholar
- Wilde F, Heufelder M, Winter K, Hendricks J, Frerich B, Schramm A, Hemprich A: The role of surgical therapy in the management of intravenous bisphosphonates-related osteonecrosis of the jaw. OOOOE. 2010Google Scholar
- Korbling M, Estrov Z: Adult stem cells for tissue repair--a new therapeutic concept?. N Engl J Med. 2003, 349: 570-582. 10.1056/NEJMra022361.View ArticlePubMedGoogle Scholar
- Schneuder RK, Puellen A, Kramann R, Raupach K, Bornemann J, Knuechel R, Perez-Buoza A, Neuss S: The osteogenic differentiation of adult bone marrow and perinatal umbilical mesenchymal stem cells and matrix remodelling in three-dimensional collagen scaffolds. Biomaterial. 2010, 31: 467-480. 10.1016/j.biomaterials.2009.09.059.View ArticleGoogle Scholar
- Ruggiero S, Gralow J, Marx RE, Hoff AO, Schubert MM, Huryn JM, Toth B, Damato K, Valero V: Practical guidelines for the prevention, diagnosis, and treatment of osteonecrosis of the jaw in patients with cancer. J Oncol Prac. 2006, 2: 7-14. 10.1200/JOP.2.1.7.View ArticleGoogle Scholar
- Gussoni E, Soneoka Y, Strickland CD, Buzney EA, Khan MK, Flint AF, Kunkel LM, Mulligan RC: Dystrophin expression in the mdx mouse restored by stem cell transplantation. Nature. 1999, 401: 309-394.Google Scholar
- Petersen BE, Bowen WC, Patrene KD, Mars WM, Sullivan AK, Murase N, Boggs SS, Greenberger JS, Goff JP: Bone marrow as a potential source of hepatic oval cells. Science. 1999, 284: 1168-1170. 10.1126/science.284.5417.1168.View ArticlePubMedGoogle Scholar
- Piepoli M, Vallisa D, Arbasi C, Cavanna L, Cerri L, Mori M, Passerini F, Tommasi L, Rossi A, Capuccci A: Bone marrow cell transplantation improves cardiac, autonomic, and functional indexes in acute anterior myocardial infarction patients (Cardiac Study). European Journal of Heart Failure.Google Scholar
- Marx RE: Pamidronate (Aredia) and zoledronate (Zometa) induced avascular necrosis of the jaw: a growing epidemic. J Oral Maxillofac Surg. 2003, 61: 1115-1117. 10.1016/S0278-2391(03)00720-1.View ArticlePubMedGoogle Scholar
- Wang J, Goodger NM, Pogrel MA: Osteonecrosis of the jawsmassociated with cancer chemotherapy. J Oral Maxillofac Surg. 2003, 61: 1104-1107. 10.1016/S0278-2391(03)00328-8.View ArticlePubMedGoogle Scholar
- Edwards BJ, Gounder M, McKoy JM, Boyd I, Farrugia M, Migliorati C, Marx R, Ruggiaero S, Dimopoulos M, Raisch DW, Singhal S, Carson K, Obadina E, Trifilio S, West D, Mehta J, Bennett CL: Bisphosphonate use and osteonecrosis of the jaw: Pharmacovigilance and reporting of this serious adverse event. Lancet Oncol. 2008, 9: 1166-10.1016/S1470-2045(08)70305-X.View ArticlePubMedGoogle Scholar
- Salamon A, Toldy E: Use of mesenchymal stem cells from adult bone marrow for injured tissue repair. Orv Hetil. 2009, 150 (27): 1259-1265. 10.1556/OH.2009.28666.View ArticlePubMedGoogle Scholar
- Connolly J, Grese R, Lipello L, Dehne R: Development of an osteogenic bone marrow preparation. J Bone Joint Surg. 1989, 71A: 684-691.Google Scholar
- Muschler G, Boehm C, Easley K: Aspiration to obtain osteoblast progenitor cells from human bone marrow: the influence of aspiration volume. J Bone Joint Surg. 1997, 79A: 1699-1707.Google Scholar
- Usami K, Mizuno H, Okada K, Narita Y, Aoki M, Kondo T, Mizuno D, Mase J, Nishiguchi H, Kagami H, Minoru U: Composite implantation of mesenchymal stem cells with endothelial progenitor cells enhances tissue-engineered bone formation. J Biomed Mater Res A. 2009, 90 (3): 730-41.View ArticlePubMedGoogle Scholar
- Bittira B, Shum-Tim D, Al-Khaldi A, Chiu RC: Mobilization and homing of bone marrow stromal cells in Myocardial infarction. Eur J Cardiotoracic Surg. 2003, 24: 393-398. 10.1016/S1010-7940(03)00325-7.View ArticleGoogle Scholar
- Prockop DJ: Marrow stromal cells as stem cells for nonhematopoietic tissues. Science. 1997, 276: 71-74. 10.1126/science.276.5309.71.View ArticlePubMedGoogle Scholar
- Asahara T, Takahashi T, Masuda H, Kalka C, Chen D, Iwaguro H, Inai Y, Silver M, Isner JM: VEGF contributes to postnatal neovascularization by mobilizing bone marrow-derived endothelial progenitor cells. EMBO J. 1999, 18: 3964-3972. 10.1093/emboj/18.14.3964.View ArticlePubMedPubMed CentralGoogle Scholar
- Orlic D, Kajstura J, Chimenti S, Bodine DM, Leri A, Anversa P: Bone marrow cells regenerate infracted myocardium. Nature. 2001, 410: 701-705. 10.1038/35070587.View ArticlePubMedGoogle Scholar
- Smart N, Riley PR: The stem cell movement. Circ Res. 2008, 102: 1155 1168Google Scholar
- Sun J, Li SH, Liu SM, Wu J, Weisel RD, Zhuo YF, Yau TM, Li Rk, Fazel SS: Improvement in cardiac function after bone marrow cell therapy is associated with an increase in myocardial inflammation. Am J Physiol Heart Circ Physiol. 2009, 296: H43-H50.View ArticlePubMedGoogle Scholar
- Kikuiri T, Kin I, Yamaza T, Akiyama K, Zhang Q, Li Y, Chen C, Chen C, Wang S, le AD, Shi S: Cell-based immunotherapy with mesenchymal stem cells cures bisphosphonate-related osteonecrosis of the jaw - like disease in mice. J Bone Miner Res. 2010, 25 (7): 1668-79. 10.1002/jbmr.37.View ArticlePubMedPubMed CentralGoogle Scholar
- Handschel J, Meyer U: Infection, vascularization, remodelling - are stem cell the answers for bone disease of the jaw?. Head & Face Medicine. 2011, 7: 5-10.1186/1746-160X-7-5.View ArticleGoogle Scholar
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