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Carboxymethyl Hyaluronan-Stabilized Nanoparticles for Anticancer Drug Delivery.

Woodman JL, Suh MS, Zhang J, Kondaveeti Y, Burgess DJ, White BA, Prestwich GD, Kuhn LT - Int J Cell Biol (2015)

Bottom Line: CMHA stabilized particles (nCaP(CMHA)) were loaded with the chemotherapy drug cis-diamminedichloroplatinum(II) (CDDP) to form nCaP(CMHA)CDDP. nCaP(CMHA)CDDP was determined to be poorly crystalline hydroxyapatite, 200 nm in diameter with a -43 mV zeta potential. nCaP(CMHA)CDDP exhibited a two-day burst release of CDDP that tapered resulting in 86% release by 7 days.Subcutaneous BT-474EMT tumors were more reproducibly inhibited by a near tumor dose of 2.8 mg/kg CDDP than a 7 mg/kg dose nCaP(CMHA)CDDP.This was likely due to a lack of distribution of nCaP(CMHA)CDDP throughout the dense tumor tissue that limited drug diffusion.

View Article: PubMed Central - PubMed

Affiliation: Department of Materials Science and Engineering, University of Connecticut, Storrs, CT 06269, USA ; Department of Reconstructive Sciences, UConn Health, Farmington, CT 06030, USA.

ABSTRACT
Carboxymethyl hyaluronic acid (CMHA) is a semisynthetic derivative of HA that is recognized by HA binding proteins but contains an additional carboxylic acid on some of the 6-hydroxyl groups of the N-acetyl glucosamine sugar units. These studies tested the ability of CMHA to stabilize the formation of calcium phosphate nanoparticles and evaluated their potential to target therapy resistant, CD44(+)/CD24(-/low) human breast cancer cells (BT-474EMT). CMHA stabilized particles (nCaP(CMHA)) were loaded with the chemotherapy drug cis-diamminedichloroplatinum(II) (CDDP) to form nCaP(CMHA)CDDP. nCaP(CMHA)CDDP was determined to be poorly crystalline hydroxyapatite, 200 nm in diameter with a -43 mV zeta potential. nCaP(CMHA)CDDP exhibited a two-day burst release of CDDP that tapered resulting in 86% release by 7 days. Surface plasmon resonance showed that nCaP(CMHA)CDDP binds to CD44, but less effectively than CMHA or hyaluronan. nCaP(CMHA-AF488) was taken up by CD44(+)/CD24(-) BT-474EMT breast cancer cells within 18 hours. nCaP(CMHA)CDDP was as cytotoxic as free CDDP against the BT-474EMT cells. Subcutaneous BT-474EMT tumors were more reproducibly inhibited by a near tumor dose of 2.8 mg/kg CDDP than a 7 mg/kg dose nCaP(CMHA)CDDP. This was likely due to a lack of distribution of nCaP(CMHA)CDDP throughout the dense tumor tissue that limited drug diffusion.

No MeSH data available.


Related in: MedlinePlus

Surface plasmon resonance sensogram depicting binding of CMHA, HA, and nCaPCMHACDDP with immobilized CD44. All data shown has been corrected for nonspecific binding to blank channels of blocked NHS-EDC. nCaPDCDDP was used as a comparable sized control, which does not have specific interactions with CD44. HA has the highest affinity for CD44, followed by CMHA then nCaPCMHACDDP.
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fig4: Surface plasmon resonance sensogram depicting binding of CMHA, HA, and nCaPCMHACDDP with immobilized CD44. All data shown has been corrected for nonspecific binding to blank channels of blocked NHS-EDC. nCaPDCDDP was used as a comparable sized control, which does not have specific interactions with CD44. HA has the highest affinity for CD44, followed by CMHA then nCaPCMHACDDP.

Mentions: Surface plasmon resonance (SPR) was performed to assess the interaction and binding of CMHA, nCaPCMHACDDP, and HA to CD44. Human CD44 chimera was immobilized on four channels of the sensor chip and two channels were amine coupled and blocked serving as a negative control for nonspecific binding. To correct for bulk shift due to the size of the nanoparticles, nCaPDCDDP was examined as a control. As expected, HA most effectively bound to CD44 with CMHA approaching, but not equaling, the binding affinity of HA (Figure 4). nCaPCMHACDDP also binds CD44, but this binding is lower than free CMHA or HA. Importantly, this binding is specific as it overcomes any bulk interactions observed with nCaPDCDDP. SPR analysis of targeted nanoparticles is challenging. SPR systems utilize complex microfluidics that normally transport solutions containing ligands or proteins of interest, but generally not solid materials such as nCaP. Of additional concern is the ability to correct for bulk shift due to the relatively large nanoparticles passing over the sensor. nCaPDCDDP served as a comparably sized, nonspecific nanoparticle control. The density of the receptor (CD44) immobilized on the chip is inherently related to the response measured; therefore, we utilized a low density of CD44 on the chip surface [42]. The highest binding observed was for HA (60 kDa), followed by CMHA (34 kDa). The chemical modification of HA to create CMHA occurs at 15–20% of the repeating 6′-OH groups of the N-acetylglucosamine residues. The interaction of CD44 and HA has low affinity but high avidity. A single HA disaccharide contains an N-acetyl-D-glucosamine and D-glucuronic acid, HA2. It has been shown that HA6 is necessary for binding CD44, but HA10 is preferred [43]. Additionally, divalent binding occurs with HA20 and larger oligomers. This likely explains the slight reduction in binding of CMHA to CD44, due to an interruption of sugar residues by the added carboxylate groups of CMHA compared to HA.


Carboxymethyl Hyaluronan-Stabilized Nanoparticles for Anticancer Drug Delivery.

Woodman JL, Suh MS, Zhang J, Kondaveeti Y, Burgess DJ, White BA, Prestwich GD, Kuhn LT - Int J Cell Biol (2015)

Surface plasmon resonance sensogram depicting binding of CMHA, HA, and nCaPCMHACDDP with immobilized CD44. All data shown has been corrected for nonspecific binding to blank channels of blocked NHS-EDC. nCaPDCDDP was used as a comparable sized control, which does not have specific interactions with CD44. HA has the highest affinity for CD44, followed by CMHA then nCaPCMHACDDP.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4581577&req=5

fig4: Surface plasmon resonance sensogram depicting binding of CMHA, HA, and nCaPCMHACDDP with immobilized CD44. All data shown has been corrected for nonspecific binding to blank channels of blocked NHS-EDC. nCaPDCDDP was used as a comparable sized control, which does not have specific interactions with CD44. HA has the highest affinity for CD44, followed by CMHA then nCaPCMHACDDP.
Mentions: Surface plasmon resonance (SPR) was performed to assess the interaction and binding of CMHA, nCaPCMHACDDP, and HA to CD44. Human CD44 chimera was immobilized on four channels of the sensor chip and two channels were amine coupled and blocked serving as a negative control for nonspecific binding. To correct for bulk shift due to the size of the nanoparticles, nCaPDCDDP was examined as a control. As expected, HA most effectively bound to CD44 with CMHA approaching, but not equaling, the binding affinity of HA (Figure 4). nCaPCMHACDDP also binds CD44, but this binding is lower than free CMHA or HA. Importantly, this binding is specific as it overcomes any bulk interactions observed with nCaPDCDDP. SPR analysis of targeted nanoparticles is challenging. SPR systems utilize complex microfluidics that normally transport solutions containing ligands or proteins of interest, but generally not solid materials such as nCaP. Of additional concern is the ability to correct for bulk shift due to the relatively large nanoparticles passing over the sensor. nCaPDCDDP served as a comparably sized, nonspecific nanoparticle control. The density of the receptor (CD44) immobilized on the chip is inherently related to the response measured; therefore, we utilized a low density of CD44 on the chip surface [42]. The highest binding observed was for HA (60 kDa), followed by CMHA (34 kDa). The chemical modification of HA to create CMHA occurs at 15–20% of the repeating 6′-OH groups of the N-acetylglucosamine residues. The interaction of CD44 and HA has low affinity but high avidity. A single HA disaccharide contains an N-acetyl-D-glucosamine and D-glucuronic acid, HA2. It has been shown that HA6 is necessary for binding CD44, but HA10 is preferred [43]. Additionally, divalent binding occurs with HA20 and larger oligomers. This likely explains the slight reduction in binding of CMHA to CD44, due to an interruption of sugar residues by the added carboxylate groups of CMHA compared to HA.

Bottom Line: CMHA stabilized particles (nCaP(CMHA)) were loaded with the chemotherapy drug cis-diamminedichloroplatinum(II) (CDDP) to form nCaP(CMHA)CDDP. nCaP(CMHA)CDDP was determined to be poorly crystalline hydroxyapatite, 200 nm in diameter with a -43 mV zeta potential. nCaP(CMHA)CDDP exhibited a two-day burst release of CDDP that tapered resulting in 86% release by 7 days.Subcutaneous BT-474EMT tumors were more reproducibly inhibited by a near tumor dose of 2.8 mg/kg CDDP than a 7 mg/kg dose nCaP(CMHA)CDDP.This was likely due to a lack of distribution of nCaP(CMHA)CDDP throughout the dense tumor tissue that limited drug diffusion.

View Article: PubMed Central - PubMed

Affiliation: Department of Materials Science and Engineering, University of Connecticut, Storrs, CT 06269, USA ; Department of Reconstructive Sciences, UConn Health, Farmington, CT 06030, USA.

ABSTRACT
Carboxymethyl hyaluronic acid (CMHA) is a semisynthetic derivative of HA that is recognized by HA binding proteins but contains an additional carboxylic acid on some of the 6-hydroxyl groups of the N-acetyl glucosamine sugar units. These studies tested the ability of CMHA to stabilize the formation of calcium phosphate nanoparticles and evaluated their potential to target therapy resistant, CD44(+)/CD24(-/low) human breast cancer cells (BT-474EMT). CMHA stabilized particles (nCaP(CMHA)) were loaded with the chemotherapy drug cis-diamminedichloroplatinum(II) (CDDP) to form nCaP(CMHA)CDDP. nCaP(CMHA)CDDP was determined to be poorly crystalline hydroxyapatite, 200 nm in diameter with a -43 mV zeta potential. nCaP(CMHA)CDDP exhibited a two-day burst release of CDDP that tapered resulting in 86% release by 7 days. Surface plasmon resonance showed that nCaP(CMHA)CDDP binds to CD44, but less effectively than CMHA or hyaluronan. nCaP(CMHA-AF488) was taken up by CD44(+)/CD24(-) BT-474EMT breast cancer cells within 18 hours. nCaP(CMHA)CDDP was as cytotoxic as free CDDP against the BT-474EMT cells. Subcutaneous BT-474EMT tumors were more reproducibly inhibited by a near tumor dose of 2.8 mg/kg CDDP than a 7 mg/kg dose nCaP(CMHA)CDDP. This was likely due to a lack of distribution of nCaP(CMHA)CDDP throughout the dense tumor tissue that limited drug diffusion.

No MeSH data available.


Related in: MedlinePlus